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Human umbilical endothelial cells (HUVECs) have a sex: characterisation of the phenotype of male and female cells

Addis, Roberta, Campesi, Ilaria, Fois, Marco, Capobianco, Giampiero, Dessole, Salvatore, Fenu, Grazia, Montella, Andrea, Cattaneo, Maria Grazia, Vicentini, Lucia M, Franconi, Flavia·2014·Biology of Sex Differences
DOI 10.1186/s13293-014-0018-2PMC PMC4273493
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Figure 1 Correlation between neonates and maternal weight (kg) stratified by sex. (A, B) Each chart contains the equation of the line, the Spearman product moment correlation coefficient and the P value.
Figure 1 Correlation between neonates and maternal weight (kg) stratified by sex. (A, B) Each chart contains the equation of the line, the Spearman product moment correlation coefficient and the P value.CC BY · PubMed Central
Figure 2 Proliferation (A) and migration of FHUVECs and MHUVECs (B, C). Linear regression analysis (A) of the proliferation of FHUVECs (▲, n = 10) and MHUVECs (▄, n = 7). *The comparison of the two slopes indicates that the slopes are different (P < 0.001). (B) Representative photographs of wound closure at 0, 9 and 24 h taken at × 4 magnification. (C) Mean of basal migratory capacity of MHUVECs (▄, n = 8) and FHUVECs (▲, n = 9). The data are expressed as median percentage of wound closure + MAD compared with the initial area (0 h). *P = 0.024.
Figure 2 Proliferation (A) and migration of FHUVECs and MHUVECs (B, C). Linear regression analysis (A) of the proliferation of FHUVECs (▲, n = 10) and MHUVECs (▄, n = 7). *The comparison of the two slopes indicates that the slopes are different (P < 0.001). (B) Representative photographs of wound closure at 0, 9 and 24 h taken at × 4 magnification. (C) Mean of basal migratory capacity of MHUVECs (▄, n = 8) and FHUVECs (▲, n = 9). The data are expressed as median percentage of wound closure + MAD compared with the initial area (0 h). *P = 0.024.CC BY · PubMed Central
Figure 3 Electron microscopic analysis of FHUVECs and MHUVECs. ( A) FHUVECs have little smooth endoplasmic reticulum (SER), abundant lysosomes (L) and a normal cytoplasmic membrane (CM). M and N indicate the mitochondria and nucleus, respectively. (B) MHUVECs have abundant autophagic vacuoles (V) in different stages of digestion and a CM with many blebs. M and N indicate the mitochondria and nucleus, respectively. (A) Magnification × 6,900, scale bars: 2 mm. (B) Magnification × 9,000, scale bars: 2 mm.
Figure 3 Electron microscopic analysis of FHUVECs and MHUVECs. ( A) FHUVECs have little smooth endoplasmic reticulum (SER), abundant lysosomes (L) and a normal cytoplasmic membrane (CM). M and N indicate the mitochondria and nucleus, respectively. (B) MHUVECs have abundant autophagic vacuoles (V) in different stages of digestion and a CM with many blebs. M and N indicate the mitochondria and nucleus, respectively. (A) Magnification × 6,900, scale bars: 2 mm. (B) Magnification × 9,000, scale bars: 2 mm.CC BY · PubMed Central
Figure 4 H 2 O 2 levels in the supernatants obtained from FHUVECs and MHUVECs. Values are expressed as medians + MAD of 15 independent samples at 72 h (*P = 0.026).
Figure 4 H 2 O 2 levels in the supernatants obtained from FHUVECs and MHUVECs. Values are expressed as medians + MAD of 15 independent samples at 72 h (*P = 0.026).CC BY · PubMed Central
Figure 5 NOS3 mRNA expression and NOS3 protein expression in FHUVECs and MHUVECs. (A) Relative NOS3 mRNA expression. The values are expressed as the means ± SEM (*P = 0.036) of four independent experiments for each sex. (B) Representative Western blot and densitometric analysis of NOS3 expression. The values are expressed as medians + MAD (*P < 0.05) of 17 independent experiments for each sex normalised to actin levels.
Figure 5 NOS3 mRNA expression and NOS3 protein expression in FHUVECs and MHUVECs. (A) Relative NOS3 mRNA expression. The values are expressed as the means ± SEM (*P = 0.036) of four independent experiments for each sex. (B) Representative Western blot and densitometric analysis of NOS3 expression. The values are expressed as medians + MAD (*P < 0.05) of 17 independent experiments for each sex normalised to actin levels.CC BY · PubMed Central
Figure 6 Beclin-1 and LC3II to LC3I ratio in FHUVECs and MHUVECs. (A) Representative Western blot and densitometric analysis of beclin-1 expression. (B) Ratio of LC3-II to LC3-I and representative Western blot. Values for each protein are expressed as medians + MAD (*P < 0.05) from at least 18 experiments and normalised to actin levels.
Figure 6 Beclin-1 and LC3II to LC3I ratio in FHUVECs and MHUVECs. (A) Representative Western blot and densitometric analysis of beclin-1 expression. (B) Ratio of LC3-II to LC3-I and representative Western blot. Values for each protein are expressed as medians + MAD (*P < 0.05) from at least 18 experiments and normalised to actin levels.CC BY · PubMed Central
Figure 7 Akt and mTOR expression levels in FHUVECs and MHUVECs. (A) Representative Western blot and densitometric analysis of Akt expression in FHUVECs and MHUVECs. The values are expressed as medians + MAD of at least six independent experiments for each sex. Akt expression was normalised to actin levels. (B) Representative Western blot and densitometric analysis of mTOR expression obtained from FHUVECs and MHUVECs. The data are the means ± SEM of at least six independent experiments for each group normalised to actin.
Figure 7 Akt and mTOR expression levels in FHUVECs and MHUVECs. (A) Representative Western blot and densitometric analysis of Akt expression in FHUVECs and MHUVECs. The values are expressed as medians + MAD of at least six independent experiments for each sex. Akt expression was normalised to actin levels. (B) Representative Western blot and densitometric analysis of mTOR expression obtained from FHUVECs and MHUVECs. The data are the means ± SEM of at least six independent experiments for each group normalised to actin.CC BY · PubMed Central
Figure 8 Protein expression of ERs, GPER and AR in FHUVECs and MHUVECs. (A, B) Representative Western blot and densitometric analysis of ER-α and ER-β expression, respectively, in FHUVECs and MHUVECs. Data are reported as the means ± SEM of at least six independent experiments for each group normalised on actin. (C, D) Representative Western blot and densitometric analysis of GPER and AR obtained from FHUVECs and MHUVECs. Data are reported as medians ± MAD for GPER30 and as means ± SEM for AR of at least four independent experiments for each group normalised on actin.
Figure 8 Protein expression of ERs, GPER and AR in FHUVECs and MHUVECs. (A, B) Representative Western blot and densitometric analysis of ER-α and ER-β expression, respectively, in FHUVECs and MHUVECs. Data are reported as the means ± SEM of at least six independent experiments for each group normalised on actin. (C, D) Representative Western blot and densitometric analysis of GPER and AR obtained from FHUVECs and MHUVECs. Data are reported as medians ± MAD for GPER30 and as means ± SEM for AR of at least four independent experiments for each group normalised on actin.CC BY · PubMed Central

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Cited by

Metadata

Authors
Addis, R., Campesi, I., Fois, M., Capobianco, G., Dessole, S., Fenu, G., Montella, A., Cattaneo, M. G., Vicentini, L. M., & Franconi, F.
Venue
Biology of Sex Differences
Publisher
Springer Science and Business Media LLC
Published
2014-12
Volume
5
Issue
1
ISSN
2042-6410
DOI
10.1186/s13293-014-0018-2
PMC
PMC4273493
Cited by
104
References
4
Language
en
Licence
creativecommons.org/licenses/by/4.0 (unspecified)

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