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MBD5 and MBD6 stabilize the BAP1 complex and promote BAP1-dependent cancer

Tsuboyama, Natsumi, Szczepanski, Aileen Patricia, Zhao, Zibo, Wang, Lu·2022·Genome Biology
DOI 10.1186/s13059-022-02776-xPMC PMC9523997
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Abstract

Background: BRCA1-associated protein 1 (BAP1) is an ubiquitin carboxy-terminal hydrolase, which forms a multi-protein complex with different epigenetic factors, such as ASXL1-3 and FOXK1/2. At the chromatin level, BAP1 catalyzes the removal of mono-ubiquitination on histone H2AK119 in collaboration with other subunits within the complex and functions as a transcriptional activator in mammalian cells. However, the crosstalk between different subunits and how these subunits impact BAP1’s function remains unclear. Results: We report the identification of the methyl-CpG-binding domain proteins 5 and 6 (MBD5 and MBD6) that bind to the C-terminal PHD fingers of the large scaffold subunits ASXL1-3 and stabilize the BAP1 complex at the chromatin. We further identify a novel Drosophila protein, the six-banded (SBA), as an ortholog of human MBD5 and MBD6, and demonstrate that the core modules of the BAP1 complex is structurally and functionally conserved from Drosophila (Calypso/ASX/SBA) to human cells (BAP1/ASXL/MBD). Dysfunction of the BAP1 complex induced by the misregulation/mutations in its subunit(s) are frequent in many human cancers. In BAP1-dependent human cancers, such as small cell lung cancer (SCLC), MBD6 tends to be a part of the predominant complex formed. Therefore, depletion of MBD6 leads to a global loss of BAP1 occupancy at the chromatin, resulting in a reduction of BAP1-dependent gene expression and tumor growth in vitro and in vivo. Conclusions: We characterize MBD5 and MBD6 as important regulators of the BAP1 complex and maintain its transcriptional landscape, shedding light on the therapeutic potential of targeting MBD5 and MBD6 in BAP1-dependent human cancers.

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Fig. 1 Endogenous MBD5 and MBD6 are stable components in the BAP1 complex. A HEK293T cells were infected by lentivirus expressing GFP, GFP-tagged MBD5, or GFP-tagged-MBD6. The protein levels of GFP, GFP-MBD5, and GFP-MBD6 were determined by western blot, n=2. B The GFP-fusion proteins were purified from HEK293T cells stably expressing GFP, GFP-MBD5, or GFP-MBD6 defined in A. The purified proteins were subjected to mass spectrometry analysis. Peptide numbers of subunits within the BAP1 complex pulled down by GFP-tagged proteins are shown. C Immunoprecipitation (IP) of endogenous BAP1 from HEK293T cells followed by immunoblot (IB) for BAP1, MBD5, and MBD6. IgG was used as a negative control, n = 2. D IP of endogenous MBD5 or MBD6 from HEK293T cells followed by IB for MBD5, MBD6, BAP1, ASXL1, and ASXL2. IgG was used as a negative control, n = 2. E Nuclear extract from HEK293T cells was subjected to size exclusion (SE) chromatography and then protein levels of BAP1, MBD5, and MBD6 were determined by western blot analysis, n = 2. F The average plots shown represent the chromatin occupancy of H3K27Ac, H3K4me1, H3K4me3, and H3K27me3 centered on MBD5 peaks (F) or MBD6 peaks (G). H Sorted and centered heatmaps generated from ChIP-seq data analyses show the occupancy of BAP1, MBD5, and MBD6 in HEK293T cells. All rows are centered on BAP1 peaks based on the ranking of signals. I The average plot shows the co-occupancy between BAP1, MBD5, and MBD6 centered on BAP1 peaks. J The representative tracks show H3K27Ac, H3K4me1, and H3K4me3 levels at BAP1, MBD5, and MBD6 occupied loci in HEK293T cells. K A Venn diagram representation of the overlapped ChIP-seq peaks between BAP1, MBD5, and MBD6
Fig. 1 Endogenous MBD5 and MBD6 are stable components in the BAP1 complex. A HEK293T cells were infected by lentivirus expressing GFP, GFP-tagged MBD5, or GFP-tagged-MBD6. The protein levels of GFP, GFP-MBD5, and GFP-MBD6 were determined by western blot, n=2. B The GFP-fusion proteins were purified from HEK293T cells stably expressing GFP, GFP-MBD5, or GFP-MBD6 defined in A. The purified proteins were subjected to mass spectrometry analysis. Peptide numbers of subunits within the BAP1 complex pulled down by GFP-tagged proteins are shown. C Immunoprecipitation (IP) of endogenous BAP1 from HEK293T cells followed by immunoblot (IB) for BAP1, MBD5, and MBD6. IgG was used as a negative control, n = 2. D IP of endogenous MBD5 or MBD6 from HEK293T cells followed by IB for MBD5, MBD6, BAP1, ASXL1, and ASXL2. IgG was used as a negative control, n = 2. E Nuclear extract from HEK293T cells was subjected to size exclusion (SE) chromatography and then protein levels of BAP1, MBD5, and MBD6 were determined by western blot analysis, n = 2. F The average plots shown represent the chromatin occupancy of H3K27Ac, H3K4me1, H3K4me3, and H3K27me3 centered on MBD5 peaks (F) or MBD6 peaks (G). H Sorted and centered heatmaps generated from ChIP-seq data analyses show the occupancy of BAP1, MBD5, and MBD6 in HEK293T cells. All rows are centered on BAP1 peaks based on the ranking of signals. I The average plot shows the co-occupancy between BAP1, MBD5, and MBD6 centered on BAP1 peaks. J The representative tracks show H3K27Ac, H3K4me1, and H3K4me3 levels at BAP1, MBD5, and MBD6 occupied loci in HEK293T cells. K A Venn diagram representation of the overlapped ChIP-seq peaks between BAP1, MBD5, and MBD6CC BY · PubMed Central
Fig. 2 The ASXL subunits link MBD5, MBD6 to BAP1 via their C-terminal PHD fingers. A IP of endogenous MBD5 or MBD6 from wild-type or BAP1 knockout HEK293T cells followed by IB for BAP1, ASXL1, and ASXL2. IgG was used as a negative control, n = 2. B IP of endogenous MBD5 or MBD6 from ASXL1/2/3 triple-knockout (TKO) HEK293T cells followed by IB for BAP1, ASXL1, ASXL2, MBD5, and MBD6. IgG was used as a negative control, n = 2. C Schematic diagram depicting the truncations of human ASXL1 protein. D HEK293T cells were transfected with plasmids expressing GFP, GFP-tagged full-length, or truncated ASXL1. The levels of GFP-tagged proteins were determined by western blot analysis, n = 2. E, F HEK293T cells were co-transfected with plasmids expressing each fragment of ASXL1 and either Halo-tagged MBD5 (E) or MBD6 (F). Then, the cells were subjected to co-IP assay using GFP-trap agarose followed by western blotting using antibodies against BAP1 and Halo-tag, n = 2. G Schematic diagram depicting the truncations of C-terminus of human ASXL1 protein. H HEK293T cells were transfected with plasmids expressing either GFP, GFP-tagged full-length, or GFP-tagged truncated C-terminus of ASXL1 and the protein levels of GFP-tagged proteins were determined by western blot analysis, n = 2. I, J HEK293T cells were co-transfected with plasmids expressing GFP-tagged truncated C-terminus of ASXL1 and either Halo-tagged MBD6 (I) or MBD5 (J). They were subjected to co-IP assay using an antibody against GFP followed by western blotting using antibodies against Halo-tag, n = 2. K CLUSTALW alignment shows the similarity between PHD fingers of ASXL1/2/3. L HEK293T cells were transfected with GFP-tagged PHD fingers of ASXL1/2/3 followed by IP of GFP and IB of MBD5, MBD6, and BAP1, n = 2. M The GFP-fusion proteins were purified from HEK293T cells transduced with GFP-tagged PHD finger of ASXL1/2/3. The purified proteins were subjected to mass spectrometry analysis. Peptide numbers of representative proteins were shown. N Schematic diagram depicting the PHD finger depletion of human ASXL1/2 proteins targeted by sgRNA. O, P IP of MBD6 from wild-type cells and ASXL1-PHD finger deleted (O) or ASXL2-PHD finger deleted (P) cells followed by IB of MBD6 and ASXL1 (O) or ASXL2 (P), n = 2
Fig. 2 The ASXL subunits link MBD5, MBD6 to BAP1 via their C-terminal PHD fingers. A IP of endogenous MBD5 or MBD6 from wild-type or BAP1 knockout HEK293T cells followed by IB for BAP1, ASXL1, and ASXL2. IgG was used as a negative control, n = 2. B IP of endogenous MBD5 or MBD6 from ASXL1/2/3 triple-knockout (TKO) HEK293T cells followed by IB for BAP1, ASXL1, ASXL2, MBD5, and MBD6. IgG was used as a negative control, n = 2. C Schematic diagram depicting the truncations of human ASXL1 protein. D HEK293T cells were transfected with plasmids expressing GFP, GFP-tagged full-length, or truncated ASXL1. The levels of GFP-tagged proteins were determined by western blot analysis, n = 2. E, F HEK293T cells were co-transfected with plasmids expressing each fragment of ASXL1 and either Halo-tagged MBD5 (E) or MBD6 (F). Then, the cells were subjected to co-IP assay using GFP-trap agarose followed by western blotting using antibodies against BAP1 and Halo-tag, n = 2. G Schematic diagram depicting the truncations of C-terminus of human ASXL1 protein. H HEK293T cells were transfected with plasmids expressing either GFP, GFP-tagged full-length, or GFP-tagged truncated C-terminus of ASXL1 and the protein levels of GFP-tagged proteins were determined by western blot analysis, n = 2. I, J HEK293T cells were co-transfected with plasmids expressing GFP-tagged truncated C-terminus of ASXL1 and either Halo-tagged MBD6 (I) or MBD5 (J). They were subjected to co-IP assay using an antibody against GFP followed by western blotting using antibodies against Halo-tag, n = 2. K CLUSTALW alignment shows the similarity between PHD fingers of ASXL1/2/3. L HEK293T cells were transfected with GFP-tagged PHD fingers of ASXL1/2/3 followed by IP of GFP and IB of MBD5, MBD6, and BAP1, n = 2. M The GFP-fusion proteins were purified from HEK293T cells transduced with GFP-tagged PHD finger of ASXL1/2/3. The purified proteins were subjected to mass spectrometry analysis. Peptide numbers of representative proteins were shown. N Schematic diagram depicting the PHD finger depletion of human ASXL1/2 proteins targeted by sgRNA. O, P IP of MBD6 from wild-type cells and ASXL1-PHD finger deleted (O) or ASXL2-PHD finger deleted (P) cells followed by IB of MBD6 and ASXL1 (O) or ASXL2 (P), n = 2CC BY · PubMed Central
Fig. 3 MBD5 and MBD6 evolutionally contributes to the stability of the BAP1 complex. A Alignment by CLUSTALW analysis shows the similarity between MBD domains of human MBD5 and MBD6. B HEK293T cells were co-transfected with plasmids expressing GFP-tagged PHD finger of ASXL1 and each Halo-tagged truncated MBD domain of MBD6. Then the cells were subjected to co-IP assay using GFP-trap agarose followed by IB for Halo-tag, n = 2. C HEK293T cells were transfected with plasmids expressing Halo-tagged MBD6-WT, MBD6-Δ12aa, or MBD6-K61A/C66A together with GFP-tagged ASXL1 followed by IP of GFP and IB for Halo-tag, n = 2. D HEK293T cells were transduced with lentivirus expressing GFP, GFP-tagged MBD6-WT, MBD6-Δ12aa, or MBD6-K61A/C66A. The protein levels of ASXL1, ASXL2, and BAP1 were determined by western blot. HSP90 was used as an internal control, n = 2. E, F HEK293T cells stably expressing GFP, GFP-tagged MBD5, or GFP-tagged MBD6 were treated with CHX (50 μg/ml) for indicated time durations. The whole cell lysate was collected at each time point, and the total protein level of BAP1 was determined by western blot, n=2 (E) and quantified by ImageJ (F). G The computational BLAST with the 12-aa from human MBD5 and MBD6 in Drosophila protein database. H Schematic diagram depicting human MBD5 and MBD6 and isoforms of Drosophila SBA. I The GFP-fusion proteins were purified from Drosophila S2 cells transduced with the GFP-tagged MBD domain of SBA. The purified proteins were subjected to mass spectrometry analysis. Peptide numbers of representative proteins pulled down by the GFP-MBD domain of SBA were shown. JDrosophila S2 cells were subjected to co-IP assay using an antibody against Calypso followed by IB for ASX and SBA, n = 2. KDrosophila S2 cells were subjected to co-IP assay using an antibody against SBA followed by IB for SBA and ASX, n = 2. LDrosophila S2 cells were subjected to co-IP assay using an antibody against ASX followed by IB for ASX and SBA, n = 2. M The western blot shows protein levels of SBA, ASX, and Calypso in Drosophila S2 cells treated with dsRNA targeting SBA. Histone H3 was used as internal control, n = 2. N Schema of the Drosophila PR-DUB complex and the human BAP1 complex
Fig. 3 MBD5 and MBD6 evolutionally contributes to the stability of the BAP1 complex. A Alignment by CLUSTALW analysis shows the similarity between MBD domains of human MBD5 and MBD6. B HEK293T cells were co-transfected with plasmids expressing GFP-tagged PHD finger of ASXL1 and each Halo-tagged truncated MBD domain of MBD6. Then the cells were subjected to co-IP assay using GFP-trap agarose followed by IB for Halo-tag, n = 2. C HEK293T cells were transfected with plasmids expressing Halo-tagged MBD6-WT, MBD6-Δ12aa, or MBD6-K61A/C66A together with GFP-tagged ASXL1 followed by IP of GFP and IB for Halo-tag, n = 2. D HEK293T cells were transduced with lentivirus expressing GFP, GFP-tagged MBD6-WT, MBD6-Δ12aa, or MBD6-K61A/C66A. The protein levels of ASXL1, ASXL2, and BAP1 were determined by western blot. HSP90 was used as an internal control, n = 2. E, F HEK293T cells stably expressing GFP, GFP-tagged MBD5, or GFP-tagged MBD6 were treated with CHX (50 μg/ml) for indicated time durations. The whole cell lysate was collected at each time point, and the total protein level of BAP1 was determined by western blot, n=2 (E) and quantified by ImageJ (F). G The computational BLAST with the 12-aa from human MBD5 and MBD6 in Drosophila protein database. H Schematic diagram depicting human MBD5 and MBD6 and isoforms of Drosophila SBA. I The GFP-fusion proteins were purified from Drosophila S2 cells transduced with the GFP-tagged MBD domain of SBA. The purified proteins were subjected to mass spectrometry analysis. Peptide numbers of representative proteins pulled down by the GFP-MBD domain of SBA were shown. JDrosophila S2 cells were subjected to co-IP assay using an antibody against Calypso followed by IB for ASX and SBA, n = 2. KDrosophila S2 cells were subjected to co-IP assay using an antibody against SBA followed by IB for SBA and ASX, n = 2. LDrosophila S2 cells were subjected to co-IP assay using an antibody against ASX followed by IB for ASX and SBA, n = 2. M The western blot shows protein levels of SBA, ASX, and Calypso in Drosophila S2 cells treated with dsRNA targeting SBA. Histone H3 was used as internal control, n = 2. N Schema of the Drosophila PR-DUB complex and the human BAP1 complexCC BY · PubMed Central
Fig. 4 MBD6 but not MBD5 is necessary for tumor cell growth. A The human SCLC cell line NCI-H1963 cells were transduced with either non-targeting CRISPR-Cas9 or two distinct CRISPR-Cas9 sgRNAs of MBD5 and MBD6. The protein levels of MBD5, MBD6, ASXL1-3, and BAP1 were determined by western blot, n=2. B Three different human SCLC cell lines were transduced with either non-targeting CRISPR gRNA, MBD5-specific gRNAs, or MBD6-specific gRNAs for 4 days. The cell numbers were determined by cell counting assay, n=3. Two-tailed unpaired Student’s t test, **P < 0.01; *P < 0.05. C The box plot shows the MBD5 and MBD6 dependency scores of different lung cancer cell types retrieved from the DepMap database. RNA-seq was performed for NCI-H1963 cells transduced with either non-targeting sgRNA, two different MBD5-specific sgRNAs, or two different MBD6-specific sgRNAs. The heatmaps (D) and the MA plot (E) show the differentially expressed genes by the depletion of MBD5 or MBD6, n = 2. F Pathway analysis by Metascape of genes that are downregulated upon MBD6 depletion in NCI-H1963 cells. G The mRNA levels of NEUROG2, DDC, FOXA2, NKX2-8, CDH7, NKX2-1, NXK2-2, and CDK18 genes, which are all critical genes for SCLC development were determined by real-time qPCR in NCI-H1963 cells transduced with non-targeting sgRNA or two different MBD6-specific sgRNAs, n=3. Two-tailed unpaired Student’s t test, **P < 0.01; *P < 0.05. H 1×106 of NCI-H510 SCLC cells transduced with either non-targeting sgRNAs or two distinct MBD6 sgRNAs that were then inoculated into the right flank of athymic nude mice, n=10 per group. Images of representative tumor tissue samples from each mouse were taken at the end of the experiment. I The tumor growth was measured using a calibrated caliper every 2–3 days. Weltch’s t-test was used for statistical analysis. J When each tumor reached 1 cm3, the mouse was euthanized and the survival probability was shown. Log-rank test was performed
Fig. 4 MBD6 but not MBD5 is necessary for tumor cell growth. A The human SCLC cell line NCI-H1963 cells were transduced with either non-targeting CRISPR-Cas9 or two distinct CRISPR-Cas9 sgRNAs of MBD5 and MBD6. The protein levels of MBD5, MBD6, ASXL1-3, and BAP1 were determined by western blot, n=2. B Three different human SCLC cell lines were transduced with either non-targeting CRISPR gRNA, MBD5-specific gRNAs, or MBD6-specific gRNAs for 4 days. The cell numbers were determined by cell counting assay, n=3. Two-tailed unpaired Student’s t test, **P < 0.01; *P < 0.05. C The box plot shows the MBD5 and MBD6 dependency scores of different lung cancer cell types retrieved from the DepMap database. RNA-seq was performed for NCI-H1963 cells transduced with either non-targeting sgRNA, two different MBD5-specific sgRNAs, or two different MBD6-specific sgRNAs. The heatmaps (D) and the MA plot (E) show the differentially expressed genes by the depletion of MBD5 or MBD6, n = 2. F Pathway analysis by Metascape of genes that are downregulated upon MBD6 depletion in NCI-H1963 cells. G The mRNA levels of NEUROG2, DDC, FOXA2, NKX2-8, CDH7, NKX2-1, NXK2-2, and CDK18 genes, which are all critical genes for SCLC development were determined by real-time qPCR in NCI-H1963 cells transduced with non-targeting sgRNA or two different MBD6-specific sgRNAs, n=3. Two-tailed unpaired Student’s t test, **P < 0.01; *P < 0.05. H 1×106 of NCI-H510 SCLC cells transduced with either non-targeting sgRNAs or two distinct MBD6 sgRNAs that were then inoculated into the right flank of athymic nude mice, n=10 per group. Images of representative tumor tissue samples from each mouse were taken at the end of the experiment. I The tumor growth was measured using a calibrated caliper every 2–3 days. Weltch’s t-test was used for statistical analysis. J When each tumor reached 1 cm3, the mouse was euthanized and the survival probability was shown. Log-rank test was performedCC BY · PubMed Central
Fig. 5 Characterization of MBD6 occupancy in SCLC cells. A The representative tracks show chromatin occupancy of MBD6, ASXL1/2/3, and BAP1 binding sites in the human SCLC cell line NCI-H1963. B The bar plot shows the feature distribution of MBD6, ASXL1/2/3, and BAP1 peaks in NCI-H1963 cells. C Sorted and centered heatmaps generated from ChIP-seq and ATAC-seq data analyses show the occupancy of MBD6, ASXL1/2/3, BAP1, H3K27Ac, H3K4me1, H3K4me3, and H3K27me3 in NCI-H1963 cells. All signals were centered on MBD6 peaks, which were classified into three clusters, based on k-means clustering according to MBD6, H3K4me1, H3K4me3, and H3K27Ac ChIP-seq. D Representative tracks showing the occupancy MBD6, BAP1, ASXL1-3, and histone marks. E Pathway analysis was performed using ChIPseeker with genes nearest to MBD6 three cluster peaks defined in C. F Motif enrichment analysis of MBD6 three clusters’ peaks defined in C from NCI-H1963 cells. G The box plot shows the log2-fold-change of H3K4me1, H3K4me3, and H3K27Ac signals versus input at MBD6/BAP1/ASXL1, MBD6/BAP1/ASXL2, or MBD6/BAP1/ASXL3 co-occupied genomic regions
Fig. 5 Characterization of MBD6 occupancy in SCLC cells. A The representative tracks show chromatin occupancy of MBD6, ASXL1/2/3, and BAP1 binding sites in the human SCLC cell line NCI-H1963. B The bar plot shows the feature distribution of MBD6, ASXL1/2/3, and BAP1 peaks in NCI-H1963 cells. C Sorted and centered heatmaps generated from ChIP-seq and ATAC-seq data analyses show the occupancy of MBD6, ASXL1/2/3, BAP1, H3K27Ac, H3K4me1, H3K4me3, and H3K27me3 in NCI-H1963 cells. All signals were centered on MBD6 peaks, which were classified into three clusters, based on k-means clustering according to MBD6, H3K4me1, H3K4me3, and H3K27Ac ChIP-seq. D Representative tracks showing the occupancy MBD6, BAP1, ASXL1-3, and histone marks. E Pathway analysis was performed using ChIPseeker with genes nearest to MBD6 three cluster peaks defined in C. F Motif enrichment analysis of MBD6 three clusters’ peaks defined in C from NCI-H1963 cells. G The box plot shows the log2-fold-change of H3K4me1, H3K4me3, and H3K27Ac signals versus input at MBD6/BAP1/ASXL1, MBD6/BAP1/ASXL2, or MBD6/BAP1/ASXL3 co-occupied genomic regionsCC BY · PubMed Central
Fig. 6 MBD6 is essential for BAP1-dependent gene expression in SCLC. The average plot (A) and representative tracks (B) show the global reduction of BAP1 in MBD6-depleted SCLC cells by two distinct sgRNAs. C Gene Set Enrichment Analysis (GSEA) of BAP1 gene signature enrichment in MBD6-depleted conditions. D The log2 fold-change heatmap shows the reduction of BAP1 peaks at MBD6 loci in cells transduced with two distinct MBD6-specific sgRNAs. E The log2 fold-change heatmap shows the change of expression levels of genes nearest to MBD6 peaks after MBD6 (left two lanes) and BAP1 (right two lanes) depletion by CRISPR, n=2. F NCI-H1963 cells were transduced with either non-targeting sgRNA or two distinct sgRNA specific to ASXL1/2/3 or BAP1. The protein levels of BAP1, ASXL1, ASXL2, and ASXL3 were determined by western blot. Tubulin was used as an internal control, n = 2. G The log2 fold-change gene expression heatmap shows BAP1/MBD6 co-targeted genes in ASXL1/2/3-depleted conditions by CRISPR knockout, n=2. H Pathway analysis by Metascape of genes that are commonly downregulated upon BAP1, MBD6, or ASXL3 depletion in NCI-H1963 cells. I The mRNA levels of NEUROG2 and CDK18 were determined by real-time qPCR in NCI-H1963 cells transduced with either non-targeting sgRNA, two different BAP1-specific sgRNAs, or two different ASXL1/2/3 sgRNAs
Fig. 6 MBD6 is essential for BAP1-dependent gene expression in SCLC. The average plot (A) and representative tracks (B) show the global reduction of BAP1 in MBD6-depleted SCLC cells by two distinct sgRNAs. C Gene Set Enrichment Analysis (GSEA) of BAP1 gene signature enrichment in MBD6-depleted conditions. D The log2 fold-change heatmap shows the reduction of BAP1 peaks at MBD6 loci in cells transduced with two distinct MBD6-specific sgRNAs. E The log2 fold-change heatmap shows the change of expression levels of genes nearest to MBD6 peaks after MBD6 (left two lanes) and BAP1 (right two lanes) depletion by CRISPR, n=2. F NCI-H1963 cells were transduced with either non-targeting sgRNA or two distinct sgRNA specific to ASXL1/2/3 or BAP1. The protein levels of BAP1, ASXL1, ASXL2, and ASXL3 were determined by western blot. Tubulin was used as an internal control, n = 2. G The log2 fold-change gene expression heatmap shows BAP1/MBD6 co-targeted genes in ASXL1/2/3-depleted conditions by CRISPR knockout, n=2. H Pathway analysis by Metascape of genes that are commonly downregulated upon BAP1, MBD6, or ASXL3 depletion in NCI-H1963 cells. I The mRNA levels of NEUROG2 and CDK18 were determined by real-time qPCR in NCI-H1963 cells transduced with either non-targeting sgRNA, two different BAP1-specific sgRNAs, or two different ASXL1/2/3 sgRNAsCC BY · PubMed Central

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Cited by

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Metadata

Authors
Tsuboyama, N., Szczepanski, A. P., Zhao, Z., & Wang, L.
Venue
Genome Biology
Publisher
Springer Science and Business Media LLC
Published
2022-09-30
Volume
23
Issue
1
ISSN
1474-760X
DOI
10.1186/s13059-022-02776-x
PMC
PMC9523997
Cited by
21
References
4
Language
en
Licence
Funders
  • American Cancer Society (RSG-22-039-01-DMC)
  • National Institute of General Medical Sciences (R35GM146979)
  • Lynn Sage Cancer Research Foundation

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CoChat. (2026). Citation check: MBD5 and MBD6 stabilize the BAP1 complex and promote BAP1-dependent cancer. https://app.cochat.ai/papers/10.1186/s13059-022-02776-x
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Tsuboyama, N., Szczepanski, A. P., Zhao, Z., & Wang, L. (2022). MBD5 and MBD6 stabilize the BAP1 complex and promote BAP1-dependent cancer. Genome Biology. https://doi.org/10.1186/s13059-022-02776-x