Fig. 7: Neat1 inhibits the expression of myogenic genes via Ezh2.

a, b ChIP-qPCR results revealing that Neat1 knockdown significantly decreased the enrichments of Ezh2 (a) and H3k27me3 (b) at the Myog, Myh4, and Tnni2 promoters. c, d ChIP-qPCR results indicating Neat1 overexpression significantly increased the enrichments of Ezh2 (c) and H3k27me3 (d) at the Myog, Myh4, and Tnni2 promoters. e Neat1 expression vector and Ezh2 siRNA fragment were co-transfected into C2C12 cells. The indicated genes expression were measured by qPCR after Neat1 expression vector and Ezh2 siRNA fragment were co-transfected 3 days post differentiation. The results showed that overexpression of Neat1 inhibits the expression of Myog, Myhc and Tnni2, but had no significant effect when co-transfected with Ezh2 siRNA fragment. f Neat1 siRNA fragment was con-transfected with Ezh2 expression vector into C2C12 cells at 5 days post differentiation, the cells were performed with Myhc immunofluorescence staining. The quantification of Myhc immunofluorescence staining results showed Neat1 knockdown alone increased Myhc protein expression but not when co-transfected with Ezh2 expression vector. g, h ChIRP-qPCR results revealed that Neat1 binds directly to the Myog, Myh4, and Tnni2 promoters but not to the Myod promoter in C2C12 cells (g) and primary myoblasts (h). Relative RNA levels were normalized to those of β-actin. All values represent the mean ± s.d. of three independent experiments. *p < 0.05, **p < 0.01, N.S. indicates not significant