Fig. 3
Silencing ZNF521 activates progenitor cell commitment and induces expression of early adipogenic factors. Silencing of ZNF521 was performed 72 h before initiation of differentiation (0 h). a Immunoblot of P16INK4 after silencing ZNF521. Immunoblots from one individual. Graph shows normalization to scrambled cells at the same time point, n = 4 independent experiments, mean ± SEM. Paired two-tailed Student′s t-test was used for comparison with scrambled cells. b Immunoblots showing cell cycle proteins. Representative immunoblots from one individual of four. c–f Expression of the early adipogenic genes. Paired two-tailed Student′s t-test was used for comparison of silenced with scrambled cells. Data represent means ± SEM *P < 0.05 and ** P < 0.01 n = 5 independent experiments. g Induction of BMP4 when ZNF521 is silenced. Data represent means ± SEM, n = 5. g–j mRNA results were first normalized to 18 S and then normalized to expression levels in the undifferentiated scrambled cells at -24 h before initiation of differentiation. h Immunoblots of BMP4, n = 4 independent experiments and i ZNF423 in whole cell lysates, n = 4 independent experiments and j in nuclear extracts, n = 3 independent experiments. g–j Paired two-tailed Student′s t-test was used for comparison of silenced with scrambled cells. *P < 0.05 and ** P < 0.01
