Fig. 5: SMAD2 promotes HFSC differentiation and apoptosis while suppressing cell proliferation by activating the NANOG/DNMT1 axis.
From: Smurf2-induced degradation of SMAD2 causes inhibition of hair follicle stem cell differentiation
A Silencing efficiency of 3 siRNAs targeting DNMT1 in HFSCs determined by RT-qPCR, with β-actin as internal reference. *p < 0.05 vs. sh-NC group. B SMAD2, NANOG, and DNMT1 protein levels in HFSCs after different treatments measured by western blot assay, with β-actin as internal reference. C Differentiation rate of HFSCs after different treatments determined by oil Red O staining. mRNA (D) and protein (E) levels of epidermal differentiation markers (K10 and involucrin) and adipogenesis markers (PPAR-γ2 and aP2), keratinocyte-specific marker (K15), and proliferation-related markers (PCNA and Ki67) in HFSCs after different treatments determined by RT-qPCR and western blot assay, with β-actin as internal reference. F Cell viability after different treatments determined by CCK-8 assay. G Cell proliferation after different treatments assessed by BrdU labeling. H The number of colonies formed in HFSCs after different treatments. I Cell apoptosis after different treatments determined by flow cytometry, *p < 0.05 vs. oe-NC group; #p < 0.05 vs. oe-SMAD2 + sh-NC group. Data are expressed as mean ± standard deviation. Data between multiple groups were compared by one-way ANOVA with Tukey’s post hoc test while data at different time points were compared by two-way ANOVA with Bonferroni post hoc test. Each cell experiment was repeated 3 times.
