Fig. 5.

PXD101 upregulates GADD45A expression through histone acetylation to mediate the activation of P38. T98G cells and LN229 cells were treated with PXD101 (5µM, 1µM), respectively. (A) The acetylation level of GADD45A was detected using IP assay with acetyl lysine antibody. (B) The protein expression of acetylated histone H3, H4, and GADD45A was detected using Western blot. T98G cells and LN229 cells were transfected with GADD45A-OE plasmid. (C) LN229 cells were transfected with pGL3-basic-GADD45A-promoter or pGL3-basic-vector plasmids. Subsequently, cells were treated with DMSO or PXD101 (1µM); then, the transcriptional activity of GADD45A was detected using dual-luciferase assay. (D) The GADD45A expression was detected using Western blot. (E) The protein expressions of GADD45A, AKT, p-AKT, P38, p-P38, JNK, p-JNK, STAT3, and p-STAT3 were analyzed using Western blot. (F) T98G cells and LN229 cells were treated with PXD101 (5µM, 1µM), respectively; the protein expressions of GADD4A, P38, and p-P38 were detected using Western blot. NC: negative control