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Increased LDL receptor by SREBP2 or SREBP2-induced lncRNA LDLR-AS promotes triglyceride accumulation in fish

Cao, Xiufei, Fang, Wei, Li, Xueshan, Wang, Xiuneng, Mai, Kangsen, Ai, Qinghui·2022·iScience
DOI 10.1016/j.isci.2022.104670PMC PMC9263516
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Figure 1 LDLR induced by PA promotes TG accumulation via the uptake of LDL (A) Lipid droplets in the liver were identified by transmission electron microscope (TEM) in fish fed with FO, SO, OO, and PO diets (Scale bars, 5 μm). The red arrow indicates lipid droplets. N, nucleus; LD, lipid droplets. (B) Lipid accumulation in fish hepatocytes was observed by Oil Red O staining after treatment with DHA, LA, OA, or PA for 12 h (Scale bars, 100 μm). (C and D) RT-qPCR and Western blot assays were used to evaluate the expressions of LDLR in the liver of fish fed with different diets. Data are presented as mean ± SEM, n = 3, one-way ANOVA, ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001. (E and F) RT-qPCR and Western blot assays were used to evaluate the expressions of LDLR in the liver of fish fed with PO diets after injection with control siRNA and si-LDLR for 48 h. Data are presented as mean ± SEM, n = 3, Student’s t-test, ∗p < 0.05, ∗∗p < 0.01. (G and H) Plasma and hepatic TG contents were tested in fish fed with PO diets after injection with control siRNA and si-LDLR for 48 h. Data are presented as mean ± SEM, n = 3, Student’s t-test, ∗∗p < 0.01. (I) Oil Red O staining was used to evaluate lipid accumulation in the liver from fish fed with PO diets after injection with control siRNA and si-LDLR for 48 h (Scale bars, 100 μm). (J) Fluorescent microscopy images were used to evaluate lipid droplets accumulation in fish hepatocytes treated with PA alone, Dil-LDL alone, or a combination of PA and Dil-LDL for 6 h (Scale bars, 25 μm). DAPI displays blue; BODIPY displays green; Dil-LDL displays red. FO, fish oil; SO, soybean oil; OO, olive oil, PO, palm oil; DHA, docosahexaenoic acid; LA, linoleic acid; OA, oleic acid; PA, palmitic acid. See also Figure S1.
Figure 1 LDLR induced by PA promotes TG accumulation via the uptake of LDL (A) Lipid droplets in the liver were identified by transmission electron microscope (TEM) in fish fed with FO, SO, OO, and PO diets (Scale bars, 5 μm). The red arrow indicates lipid droplets. N, nucleus; LD, lipid droplets. (B) Lipid accumulation in fish hepatocytes was observed by Oil Red O staining after treatment with DHA, LA, OA, or PA for 12 h (Scale bars, 100 μm). (C and D) RT-qPCR and Western blot assays were used to evaluate the expressions of LDLR in the liver of fish fed with different diets. Data are presented as mean ± SEM, n = 3, one-way ANOVA, ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001. (E and F) RT-qPCR and Western blot assays were used to evaluate the expressions of LDLR in the liver of fish fed with PO diets after injection with control siRNA and si-LDLR for 48 h. Data are presented as mean ± SEM, n = 3, Student’s t-test, ∗p < 0.05, ∗∗p < 0.01. (G and H) Plasma and hepatic TG contents were tested in fish fed with PO diets after injection with control siRNA and si-LDLR for 48 h. Data are presented as mean ± SEM, n = 3, Student’s t-test, ∗∗p < 0.01. (I) Oil Red O staining was used to evaluate lipid accumulation in the liver from fish fed with PO diets after injection with control siRNA and si-LDLR for 48 h (Scale bars, 100 μm). (J) Fluorescent microscopy images were used to evaluate lipid droplets accumulation in fish hepatocytes treated with PA alone, Dil-LDL alone, or a combination of PA and Dil-LDL for 6 h (Scale bars, 25 μm). DAPI displays blue; BODIPY displays green; Dil-LDL displays red. FO, fish oil; SO, soybean oil; OO, olive oil, PO, palm oil; DHA, docosahexaenoic acid; LA, linoleic acid; OA, oleic acid; PA, palmitic acid. See also Figure S1.CC BY-NC-ND · PubMed Central
Figure 2 SREBP2 activated by TNFα increases the transcription of LDLR (A) LDLR promoter activity was analyzed by dual-luciferase reporter after co-transfection of LDLR promoter with PCS2+ vector or 10 transcription factors involved in lipid metabolism in HEK293T cells. Data are presented as mean ± SEM, n = 4, one-way ANOVA, ∗∗p < 0.01, ∗∗∗p < 0.001. (B) LDLR promoter activity was analyzed by dual-luciferase reporter after co-transfection of LDLR promoter with PCS2+ or PCS2+SREBP2 vectors in different doses (range: 0–600 ng/μL) in HEK293T cells. Data are presented as mean ± SEM, n = 3, one-way ANOVA, ∗p < 0.05, ∗∗∗p < 0.001. (C) The schematic diagram shows that two predicted SREBP2-binding sites (a and b) contained at LDLR promoter were mutated. (D) LDLR promoter activity was analyzed by dual-luciferase reporter after co-transfection of wt/mut LDLR promoters with PCS2+ or PCS2+SREBP2 vectors in HEK293T cells. Data are presented as mean ± SEM, n = 4, one-way ANOVA, ∗∗∗p < 0.001. (E and F) The expression levels of SREBP2 mRNA and protein were analyzed by RT-qPCR and Western blot in the liver of fish fed with FO, SO, OO, and PO diets. Data are presented as mean ± SEM, n = 3, one-way ANOVA, ∗p < 0.05, ∗∗p < 0.01. (G and H) SREBP2 and LDLR mRNA expressions were detected by RT-qPCR in fish hepatocytes interfered with control siRNA or si-SREBP2 for 24 h after treatment with PA for 12 h. Data are presented as mean ± SEM, n = 3, one-way ANOVA, ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001. (I and J) Plasma and liver TNFα contents were detected in fish fed with FO, SO, OO, and PO diets. Data are presented as mean ± SEM, n = 3, one-way ANOVA, ∗p < 0.05, ∗∗p < 0.01. (K–O) Genes (TNFα, IL-1β, MMP9, COX-2, and VCAM-1) involved in inflammatory response were analyzed by RT-qPCR in the liver of fish fed with FO, SO, OO, and PO diets. Data are presented as mean ± SEM, n = 3, one-way ANOVA, ∗p < 0.05, ∗∗p < 0.01, ns = not significant. (P) Genes (TNFα, IL-6, NF-κB, MMP9, VCAM-1, COX-2, SREBP2, and LDLR) were analyzed after fish hepatocytes were treated with TNFα for 24 h. Data are presented as mean ± SEM, n = 3, Student’s t-test, ∗p < 0.05, ∗∗p < 0.01, ns = not significant. (Q and R) Western blot analysis of SREBP2 in nucleus and LDLR protein expressions in fish hepatocytes treated with TNFα for 24 h. Data are presented as mean ± SEM, n = 3, Student’s t-test, ∗p < 0.05. (S) TG content in fish hepatocytes was analyzed after TNFα treatment for 24 h. Data are presented as mean ± SEM, n = 3, Student’s t-test, ∗p < 0.05. FO, fish oil; SO, soybean oil; OO, olive oil, PO, palm oil; PA, palmitic acid. See also Figure S2.
Figure 2 SREBP2 activated by TNFα increases the transcription of LDLR (A) LDLR promoter activity was analyzed by dual-luciferase reporter after co-transfection of LDLR promoter with PCS2+ vector or 10 transcription factors involved in lipid metabolism in HEK293T cells. Data are presented as mean ± SEM, n = 4, one-way ANOVA, ∗∗p < 0.01, ∗∗∗p < 0.001. (B) LDLR promoter activity was analyzed by dual-luciferase reporter after co-transfection of LDLR promoter with PCS2+ or PCS2+SREBP2 vectors in different doses (range: 0–600 ng/μL) in HEK293T cells. Data are presented as mean ± SEM, n = 3, one-way ANOVA, ∗p < 0.05, ∗∗∗p < 0.001. (C) The schematic diagram shows that two predicted SREBP2-binding sites (a and b) contained at LDLR promoter were mutated. (D) LDLR promoter activity was analyzed by dual-luciferase reporter after co-transfection of wt/mut LDLR promoters with PCS2+ or PCS2+SREBP2 vectors in HEK293T cells. Data are presented as mean ± SEM, n = 4, one-way ANOVA, ∗∗∗p < 0.001. (E and F) The expression levels of SREBP2 mRNA and protein were analyzed by RT-qPCR and Western blot in the liver of fish fed with FO, SO, OO, and PO diets. Data are presented as mean ± SEM, n = 3, one-way ANOVA, ∗p < 0.05, ∗∗p < 0.01. (G and H) SREBP2 and LDLR mRNA expressions were detected by RT-qPCR in fish hepatocytes interfered with control siRNA or si-SREBP2 for 24 h after treatment with PA for 12 h. Data are presented as mean ± SEM, n = 3, one-way ANOVA, ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001. (I and J) Plasma and liver TNFα contents were detected in fish fed with FO, SO, OO, and PO diets. Data are presented as mean ± SEM, n = 3, one-way ANOVA, ∗p < 0.05, ∗∗p < 0.01. (K–O) Genes (TNFα, IL-1β, MMP9, COX-2, and VCAM-1) involved in inflammatory response were analyzed by RT-qPCR in the liver of fish fed with FO, SO, OO, and PO diets. Data are presented as mean ± SEM, n = 3, one-way ANOVA, ∗p < 0.05, ∗∗p < 0.01, ns = not significant. (P) Genes (TNFα, IL-6, NF-κB, MMP9, VCAM-1, COX-2, SREBP2, and LDLR) were analyzed after fish hepatocytes were treated with TNFα for 24 h. Data are presented as mean ± SEM, n = 3, Student’s t-test, ∗p < 0.05, ∗∗p < 0.01, ns = not significant. (Q and R) Western blot analysis of SREBP2 in nucleus and LDLR protein expressions in fish hepatocytes treated with TNFα for 24 h. Data are presented as mean ± SEM, n = 3, Student’s t-test, ∗p < 0.05. (S) TG content in fish hepatocytes was analyzed after TNFα treatment for 24 h. Data are presented as mean ± SEM, n = 3, Student’s t-test, ∗p < 0.05. FO, fish oil; SO, soybean oil; OO, olive oil, PO, palm oil; PA, palmitic acid. See also Figure S2.CC BY-NC-ND · PubMed Central
Figure 3 Screening and identification of lncRNA LDLR-AS associated with LDLR (A and B) Venn diagram was used for identifying novel lncRNAs and mRNAs expressed in the liver of fish fed with FO, SO, OO, and PO diets. (C) The schematic diagram of positional relationship between LDLR-AS and LDLR on the chromosome of large yellow croaker. “+” on the left side of strand represents the positive strand; “-” represents the negative strand. Red arrows indicate transcription direction, and two blocks with colors (light blue and yellow) represent the transcripts of LDLR and LDLR-AS, respectively. (D) The localization of LDLR-AS in fish hepatocytes was evaluated by nuclear/cytosolic fractionation, followed by RT-qPCR analysis. U6 and GAPDH were used as internal references of nuclear and cytoplasmic RNA, respectively. Data are presented as mean ± SEM, n = 3, Student’s t-test, ∗∗p < 0.01. (E) The subcellular distribution of Cy3-tagged LDLR-AS was visualized by RNA Fluorescent in situ hybridization (FISH) in fish hepatocytes (Scale bars, 10 μm). Nuclear were stained by DAPI. (F) Correlation between LDLR-AS and LDLR FPKM expression from transcriptome sequencing in the liver from fish fed with FO, SO, OO, and PO diets. The result was analyzed using Pearson correlation analysis (n = 12, Pearson correlation analysis, p < 0.05). (G and H) The tissue distribution of LDLR and LDLR-AS in large yellow croaker was analyzed by RT-qPCR. Data are presented as mean ± SEM, n = 3, one-way ANOVA, ∗∗p < 0.01, ∗∗∗p < 0.001. (I) RT-qPCR results of the expression of LDLR-AS in the liver of fish fed with FO, SO, OO, and PO diets. Data are presented as mean ± SEM, n = 3, one-way ANOVA, ∗p < 0.05, ∗∗p < 0.01. FO, fish oil; SO, soybean oil; OO, olive oil, PO, palm oil. See also Figure S3.
Figure 3 Screening and identification of lncRNA LDLR-AS associated with LDLR (A and B) Venn diagram was used for identifying novel lncRNAs and mRNAs expressed in the liver of fish fed with FO, SO, OO, and PO diets. (C) The schematic diagram of positional relationship between LDLR-AS and LDLR on the chromosome of large yellow croaker. “+” on the left side of strand represents the positive strand; “-” represents the negative strand. Red arrows indicate transcription direction, and two blocks with colors (light blue and yellow) represent the transcripts of LDLR and LDLR-AS, respectively. (D) The localization of LDLR-AS in fish hepatocytes was evaluated by nuclear/cytosolic fractionation, followed by RT-qPCR analysis. U6 and GAPDH were used as internal references of nuclear and cytoplasmic RNA, respectively. Data are presented as mean ± SEM, n = 3, Student’s t-test, ∗∗p < 0.01. (E) The subcellular distribution of Cy3-tagged LDLR-AS was visualized by RNA Fluorescent in situ hybridization (FISH) in fish hepatocytes (Scale bars, 10 μm). Nuclear were stained by DAPI. (F) Correlation between LDLR-AS and LDLR FPKM expression from transcriptome sequencing in the liver from fish fed with FO, SO, OO, and PO diets. The result was analyzed using Pearson correlation analysis (n = 12, Pearson correlation analysis, p < 0.05). (G and H) The tissue distribution of LDLR and LDLR-AS in large yellow croaker was analyzed by RT-qPCR. Data are presented as mean ± SEM, n = 3, one-way ANOVA, ∗∗p < 0.01, ∗∗∗p < 0.001. (I) RT-qPCR results of the expression of LDLR-AS in the liver of fish fed with FO, SO, OO, and PO diets. Data are presented as mean ± SEM, n = 3, one-way ANOVA, ∗p < 0.05, ∗∗p < 0.01. FO, fish oil; SO, soybean oil; OO, olive oil, PO, palm oil. See also Figure S3.CC BY-NC-ND · PubMed Central
Figure 4 SREBP2 promotes the transcriptional level of LDLR-AS (A) LDLR-AS expression was detected by RT-qPCR in fish hepatocytes interfered with control siRNA or si-SREBP2 for 24 h after treatment with PA for 12 h. Data are presented as mean ± SEM, n = 3, one-way ANOVA, ∗p < 0.05. (B) LDLR-AS promoter activity was analyzed by dual-luciferase reporter after co-transfection of LDLR-AS promoter with PCS2+ or PCS2-SREBP2 vectors in HEK293T cells. Data are presented as mean ± SEM, n = 4, Student’s t-test, ∗∗p < 0.01. (C) LDLR-AS promoter activity was analyzed by dual-luciferase reporter after co-transfection of four truncated LDLR-AS promoters with PCS2+ or PCS2-SREBP2 vectors in HEK293T cells. Data are presented as mean ± SEM, n = 4, one-way ANOVA. (D) The schematic diagram shows that predicted SREBP2-binding site contained at −1273 ∼ −862 bp of LDLR-AS promoter was mutated. (E) The activities of wt/mut LDLR-AS promoter were analyzed by dual-luciferase reporter after co-transfection of wt/mut LDLR-AS promoters with PCS2+ or PCS2-SREBP2 vectors in HEK293T cells. Data are presented as mean ± SEM, n = 4, one-way ANOVA, ∗∗∗p < 0.001. (F) ChIP-PCR assay of SREBP2-binding region at −1273 ∼ - 862 bp from LDLR-AS promoter using IgG or Flag antibody.
Figure 4 SREBP2 promotes the transcriptional level of LDLR-AS (A) LDLR-AS expression was detected by RT-qPCR in fish hepatocytes interfered with control siRNA or si-SREBP2 for 24 h after treatment with PA for 12 h. Data are presented as mean ± SEM, n = 3, one-way ANOVA, ∗p < 0.05. (B) LDLR-AS promoter activity was analyzed by dual-luciferase reporter after co-transfection of LDLR-AS promoter with PCS2+ or PCS2-SREBP2 vectors in HEK293T cells. Data are presented as mean ± SEM, n = 4, Student’s t-test, ∗∗p < 0.01. (C) LDLR-AS promoter activity was analyzed by dual-luciferase reporter after co-transfection of four truncated LDLR-AS promoters with PCS2+ or PCS2-SREBP2 vectors in HEK293T cells. Data are presented as mean ± SEM, n = 4, one-way ANOVA. (D) The schematic diagram shows that predicted SREBP2-binding site contained at −1273 ∼ −862 bp of LDLR-AS promoter was mutated. (E) The activities of wt/mut LDLR-AS promoter were analyzed by dual-luciferase reporter after co-transfection of wt/mut LDLR-AS promoters with PCS2+ or PCS2-SREBP2 vectors in HEK293T cells. Data are presented as mean ± SEM, n = 4, one-way ANOVA, ∗∗∗p < 0.001. (F) ChIP-PCR assay of SREBP2-binding region at −1273 ∼ - 862 bp from LDLR-AS promoter using IgG or Flag antibody.CC BY-NC-ND · PubMed Central
Figure 5 LDLR-AS affects hepatic triglyceride content by regulating LDLR expression (A) Overexpression of LDLR-AS was detected by RT-qPCR after pcDNA3.1 or pcDNA3.1-LDLR-AS plasmids were electroporated into fish hepatocytes for 48 h pcDNA3.1 plasmid was used as a negative control. Data are presented as mean ± SEM, n = 3, Student’s t-test, ∗∗∗p < 0.001. (B and C) RT-qPCR and Western blot assays were used to evaluate the expressions of LDLR in fish hepatocytes after LDLR-AS overexpression. Data are presented as mean ± SEM, n = 3, Student’s t-test, ∗p < 0.05. (D) TG content in fish hepatocytes was analyzed after LDLR-AS overexpression. Data are presented as mean ± SEM, n = 3, Student’s t-test, ∗p < 0.05. (E) LDLR-AS expression was knocked down in the PO diets group by injecting control siRNA (NC) or LDLR-AS ASO3 (ASO3). Data are presented as mean ± SEM, n = 3, Student’s t-test, ∗p < 0.05. (F and G) The expression levels of LDLR mRNA and protein were analyzed by RT-qPCR and Western blot after LDLR-AS knockdown. Data are presented as mean ± SEM, n = 3, Student’s t-test, ∗p < 0.05, ∗∗p < 0.01. (H and I) Plasma and liver TG levels were detected after LDLR-AS knockdown. Data are presented as mean ± SEM, n = 3, Student’s t-test, ∗p < 0.05. (J) Lipid accumulation in the liver was observed by Oil Red O staining after LDLR-AS knockdown (Scale bars, 100 μm). See also Figure S4.
Figure 5 LDLR-AS affects hepatic triglyceride content by regulating LDLR expression (A) Overexpression of LDLR-AS was detected by RT-qPCR after pcDNA3.1 or pcDNA3.1-LDLR-AS plasmids were electroporated into fish hepatocytes for 48 h pcDNA3.1 plasmid was used as a negative control. Data are presented as mean ± SEM, n = 3, Student’s t-test, ∗∗∗p < 0.001. (B and C) RT-qPCR and Western blot assays were used to evaluate the expressions of LDLR in fish hepatocytes after LDLR-AS overexpression. Data are presented as mean ± SEM, n = 3, Student’s t-test, ∗p < 0.05. (D) TG content in fish hepatocytes was analyzed after LDLR-AS overexpression. Data are presented as mean ± SEM, n = 3, Student’s t-test, ∗p < 0.05. (E) LDLR-AS expression was knocked down in the PO diets group by injecting control siRNA (NC) or LDLR-AS ASO3 (ASO3). Data are presented as mean ± SEM, n = 3, Student’s t-test, ∗p < 0.05. (F and G) The expression levels of LDLR mRNA and protein were analyzed by RT-qPCR and Western blot after LDLR-AS knockdown. Data are presented as mean ± SEM, n = 3, Student’s t-test, ∗p < 0.05, ∗∗p < 0.01. (H and I) Plasma and liver TG levels were detected after LDLR-AS knockdown. Data are presented as mean ± SEM, n = 3, Student’s t-test, ∗p < 0.05. (J) Lipid accumulation in the liver was observed by Oil Red O staining after LDLR-AS knockdown (Scale bars, 100 μm). See also Figure S4.CC BY-NC-ND · PubMed Central
Figure 6 LDLR-AS interacts with hnRNPR to stabilize LDLR mRNA (A) Silver staining of proteins pulled down with biotinylated LDLR-AS RNA and a negative control with antisense sequences. Blue triangle denotes the band identified as hnRNPR by mass spectrometry. (B) Interaction profiles for LDLR-AS interaction with hnRNPR were predicted using the catRAPID website. (C) ChIP-qPCR assay was performed to measure the binding ability of hnRNPR to LDLR-AS. Data are presented as mean ± SEM, n = 3, Student’s t-test, ∗∗∗p < 0.001. (D) RT-qPCR results of hnRNPR knockdown in fish hepatocytes transfected with si-hnRNPR#1/2/3 for 24 h. Data are presented as mean ± SEM, n = 3, one-way ANOVA, ∗p < 0.05. (E) Western blot of LDLR and hnRNPR protein in fish hepatocytes transfected with si-hnRNPR#1 for 48 h. Data are presented as mean ± SEM, n = 3, one-way ANOVA, ∗p < 0.05, ∗∗p < 0.01. (F) RT-qPCR results of LDLR-AS expression in fish hepatocytes transfected with si-hnRNPR#1/2/3 for 24 h. Data are presented as mean ± SEM, n = 3, one-way ANOVA, ns = not significant. (G) RT-qPCR analysis was used to evaluate the stability of LDLR mRNA after fish hepatocytes were transfected with si-hnRNPR#1 for 24 h, then further exposed to 5 μg/mL actinomycin D for 24 h. Data are presented as mean ± SEM, n = 3, Student’s t-test, ∗∗p < 0.01. (H) Interaction profiles for LDLR at the 5′-UTR interaction with hnRNPR were predicted using the catRAPID website. (I) The luciferase activity of 5′-UTR of LDLR mRNA was detected in HEK293T cells transfected pGL3-Basic, pGL3-5′UTR, or PCS2+hnRNPR vectors. Data are presented as mean ± SEM, n = 3, one-way ANOVA, ∗∗p < 0.01, ∗∗∗p < 0.001. (J) ChIP-qPCR assay was performed to measure the binding ability of hnRNPR to the 5′-UTR of LDLR. Data are presented as mean ± SEM, n = 3, Student’s t-test, ∗∗p < 0.01.
Figure 6 LDLR-AS interacts with hnRNPR to stabilize LDLR mRNA (A) Silver staining of proteins pulled down with biotinylated LDLR-AS RNA and a negative control with antisense sequences. Blue triangle denotes the band identified as hnRNPR by mass spectrometry. (B) Interaction profiles for LDLR-AS interaction with hnRNPR were predicted using the catRAPID website. (C) ChIP-qPCR assay was performed to measure the binding ability of hnRNPR to LDLR-AS. Data are presented as mean ± SEM, n = 3, Student’s t-test, ∗∗∗p < 0.001. (D) RT-qPCR results of hnRNPR knockdown in fish hepatocytes transfected with si-hnRNPR#1/2/3 for 24 h. Data are presented as mean ± SEM, n = 3, one-way ANOVA, ∗p < 0.05. (E) Western blot of LDLR and hnRNPR protein in fish hepatocytes transfected with si-hnRNPR#1 for 48 h. Data are presented as mean ± SEM, n = 3, one-way ANOVA, ∗p < 0.05, ∗∗p < 0.01. (F) RT-qPCR results of LDLR-AS expression in fish hepatocytes transfected with si-hnRNPR#1/2/3 for 24 h. Data are presented as mean ± SEM, n = 3, one-way ANOVA, ns = not significant. (G) RT-qPCR analysis was used to evaluate the stability of LDLR mRNA after fish hepatocytes were transfected with si-hnRNPR#1 for 24 h, then further exposed to 5 μg/mL actinomycin D for 24 h. Data are presented as mean ± SEM, n = 3, Student’s t-test, ∗∗p < 0.01. (H) Interaction profiles for LDLR at the 5′-UTR interaction with hnRNPR were predicted using the catRAPID website. (I) The luciferase activity of 5′-UTR of LDLR mRNA was detected in HEK293T cells transfected pGL3-Basic, pGL3-5′UTR, or PCS2+hnRNPR vectors. Data are presented as mean ± SEM, n = 3, one-way ANOVA, ∗∗p < 0.01, ∗∗∗p < 0.001. (J) ChIP-qPCR assay was performed to measure the binding ability of hnRNPR to the 5′-UTR of LDLR. Data are presented as mean ± SEM, n = 3, Student’s t-test, ∗∗p < 0.01.CC BY-NC-ND · PubMed Central

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Cited by

Sorted by citations

Metadata

Authors
Cao, X., Fang, W., Li, X., Wang, X., Mai, K., & Ai, Q.
Venue
iScience
Publisher
Elsevier BV
Published
2022-07
Volume
25
Issue
7
Pages
104670
ISSN
2589-0042
DOI
10.1016/j.isci.2022.104670
PMC
PMC9263516
Cited by
32
References
0
Language
en
Licence
Funders
  • National Natural Science Foundation of China (31830103)
  • China National Funds for Distinguished Young Scientists (31525024, CS31117200001)
  • Agriculture Research System of China (CARS47-11)

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