Figure 5. LHX9 inhibited glycolysis by down-regulating PGK1 expression. (A) Interfering with LHX9 expression up-regulated the mRNA level of PGK1. (B) Overexpression of LHX9 inhibited the induction of PGK1 by knockdown of p53. (C) Left: the schematic illustration of the PKG1 promoter with labeled p53 and LHX9 binding sites. Right: Chromosomal immunoprecipitation assay demonstrated that p53 and LHX9 bound to the PGK1 promoter. (D) Chromosomal immunoprecipitation assay demonstrated that down-regulating LHX9 expression inhibited the binding of p53 to the PGK1 promoter, and vice versa. (E, F) Soft agar assay demonstrated that down-regulating the expression of PGK1 abolished the increase in colony formation caused by down-regulation of LHX9. (G) Interfering with LHX9 expression increased the lactic acid content. This increase could be suppressed by interfering with PGK1. (H, I) The effects of P53 restoration on the anchorage-independent growth of SK-N-SH were examined. (J) The effects of P53 restoration on the PGK1 expression of SK-N-SH were examined. ##, P<0.01; **, P<0.01.