Fig 2: FGF7 is downregulated in cardiomyocytes upon MI and oxygen-glucose deprivation (OGD). (A–C) Representative images of immunofluorescence staining of (A) FGF7 (red), c-TNT (green) and DAPI (blue), (B) FGF7 (red), vimentin (green) and DAPI (blue) and (C) FGF7 (red), CD34 (green) and DAPI (blue) in the infarct zone of mouse hearts in sham group and 7 days after MI. c-TNT: cardiomyocytes marker. Vimentin: fibroblasts marker. CD34: endothelial cells marker. Scale bar, 50 µm. (D) Western blotting analysis of FGF7 expression in adult cardiomyocytes (ACMs) and adult cardiac fibroblasts (ACFs) in sham or MI mice. n = 4 in per group. (E) Cell lysates of NRCMs were used to detect the FGF7, Bax, Bcl-2 as well as c-Cas-3 protein levels by Western blot and quantified by Image J software. c-Cas-3: cleaved-Caspase-3. n = 4 in per group. All values are presented as mean ± SD. N.S: non-significant, *P < 0.05, ***P < 0.001. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)

Fig 3: MBD3 knockdown partially reversed the upregulation of MBD3 and cell differentiation induced by the removal of PKC inhibitor.A MBD3 RNA expression increased in mES at passage 5 when the PKC inhibitor was removed for 48 h and was partially reversed by MBD3 knockdown, with the PKCi and PKCi removal plus shNC transfection groups used as controls. B MBD3 and ß-actin protein levels were evaluated by Western blot (upper panel). Quantitative density analysis showed that MBD3 protein levels increased after PKC inhibitor was removed, which was partially reversed by MBD3 knockdown (lower panel). C PKC inhibitor removal decreased mRNA levels of pluripotency genes NANOG, OCT4, and SOX2 and naïve-state markers KLF4, FGF4, REX1, and NROB1, whereas MBD3 knockdown partially increased the mRNA levels of pluripotency gene, NANOG, SOX2, KLF4, and NROB1. PKC inhibitor removal increased mRNA levels of primed-state markers T and FGF5 and the mesoderm gene cTnT, whereas MBD3 knockdown partially decreased mRNA levels of T, FGF5, and cTnT. D NANOG, KLF4, FGF5, cTnT, and ß-actin levels were detected by Western blot (left panel). Quantitative density analysis showed that NANOG and KLF4 protein levels decreased and FGF5 and cTnT protein levels increased after PKC inhibitor removal, whereas NANOG and KLF4 protein levels partially increased, but FGF5 and cTnT levels partially decreased after MBD3 knockdown (right panel). E After PKC inhibitor removal, mES differentiated and did not show AP staining, whereas MBD3 knockdown partially recovered mES morphology and restored AP staining. Scale bar, 200 µm (upper panel). PKC inhibitor removal decreased the total number of AP-positive colonies, which was partially reversed by MBD3 knockdown (left panel). MBD3 knockdown partially reversed the decrease in the percentage of mixed and undifferentiated colonies and the increase in the percentage of differentiated colonies induced by PKC inhibitor removal (right panel). Data were shown as mean ± SD (n = 3). The letters a, b, and c indicated significant differences among groups (P < 0.05).