Fig. 6: HEP14 intensified the anti-fibrotic and pro-angiogenic effects of h-hADSCs via activating PKC-ERK1/2-MMP1 and -PDGFD signal cascades.

A Expression and quantification of PKC, pPKC, ERK1/2 and ERK1/2 in h-hADSCs and c-hADSCs by Western blot. B Expression and quantification of MMP1, PDGFD and PDGFRβ in HEP14-treated hADSCs and vehicle-treated-hADSCs as control by Western blot. C Representative images of IHC-stained ovarian sections and quantification with pPKC, pERK1/2 and MMP1 in four groups of normal control (CTL), vehicle-treated (POI)-, h-hADSCs-treated (h-hADSCs), and h-hADSCs/HEP14-treated (h-hADSCs/HEP14) POI ovaries in POI mice. D Representative images of ovarian sections co-stained for PDGFD and PDGFRβ, along with quantification of fluorescence intensity in four groups of POI ovaries. E Representative images of IHC-stained ovarian sections and quantification with pPKC, pERK1/2 and MMP1 in three groups of control aging-, h-hADSCs-treated (h-hADSCs)-, and h-hADSCs/HEP14-treated (h-hADSCs/HEP14) aging ovaries in aging mice. F Representative images of ovarian sections co-stained for PDGFD and PDGFRβ, along with quantification of fluorescence intensity in three groups of aging ovaries. G The level of secreted MMP-1 in the various conditioned media, including c-hADSCs-CM (CTL), h-hADSCs-CM (HEP14), h-hADSCs+PKCi-CM (HEP14+Bis I) and h-hADSCs+ERKi-CM (HEP14+PD98059) by ELISA assay. H, I Tube formation of HUVECs and quantification with various conditioned media described above, assessed by tube-formation assay, with confirmation by IF staining for CD31 and quantification of the tube length and numbers of branching points. Data are presented as means ± SEM (n = 3). P values were calculated by one-way ANOVA, followed by Tukey-Kramer post hoc test. Changes were considered statistically significant when P < 0.05. Scale bar = 100 μm.