Fig. 5
Sex differences in the activities and expression of ECM proteases of hPSC-derived smooth muscle progenitor cells (pSMCs) (a, b). Zymographic evaluation of MMP activities in concentrated condition media of male pSMCs (a) and female pSMCs (b) cocultured with different concentrations of 17β-estradiol. Zymograms are representative of three separate experiments. MMP-2 appeared as an active isoform (64/62 kDa) and a latent isoform (pro-MMP-2, 72 kDa), while MPP-1 appeared as an active isoform (43 kDa) in the gels. Graphs show the densitometric data of active-MMP-2 and/or pro-MMP-2 activities. c qRT-PCR analysis for the gene expression of MMP-1 in the male and female pSMCs at baseline (no E2 stimulation). d, e Western blot analysis of TIMP-1 protein in the concentrated supernatant from male and female pSMCs. Blots show changes in TIMP-1 protein in the concentrated condition media from male HuF1-pSMCs and female HuF5-pSMCs cocultured with or without E2. Graphs show the densitometric analysis of TIMP-1 protein expression in each group respectively. Results were normalized with internal control GAPDH. Data shown represent the mean ± SD from three independent experiments, each performed in triplicate (a, b, d, e) or duplicate (c). *p < 0.05, vs untreated pSMCs. # p < 0.05, male pSMCs vs female pSMCs. F female, M male, MMP matrix metalloproteinase, TIMP tissue inhibitor of metalloproteinase