Figure 7

Knockdown of HDAC3 or SMRT enhances NPC generation. (A) Knockdown efficiency of shHDAC1 or shHDAC2, shHDAC3 was assessed by real-time PCR. The error bars indicate SEM. **P <0.01; ***P <0.001. (B) Neurosphere formation of H9-shHDAC1 and H9-shHDAC2 cells as well as H9-shHDAC3 cells were analyzed on day 18. Scale bar, 200 μm. Black arrows represent typical neurospheres, and red arrowheads represent cell debris. (C) Co-IP of endogenous HDAC3 and SMRT from H9 cell lysate was performed with HDAC3 or SMRT antibodies. (D) Knockdown efficiency of SMRT was assessed by real-time PCR. (E, F) On day 18 of differentiation, neurospheres were formed in shNC, shHDAC3 and shSMRT H9 cells with or without MGCD treatment for seven days. Black arrows represent typical neurospheres, and red arrowheads represent cell debris. (G, H) On day 7 of differentiation, real-time PCR was performed to determine the mRNA levels of neuroectodermal genes (PAX6, SOX2 and NESTIN) and endoderm gene (GATA6) in shHDAC3 and shSMRT H9 cells with or without MGCD treatment for seven days. Scale bars, 200 μm. The error bars indicate SEM, *P <0.05; **P <0.01; ***P <0.001; n >/=3. Co-IP, co-immunoprecipitation; HDAC, histone deacetylase; NPC, neural progenitor cell; SEM, standard error of the mean.