Fig. 6

AS-tDR-007333 activates HSPB1 to regulate H3K4me1 and H3K27ac levels in MED29 promoter region. A AS-tDR-007333 up-regulated HSPB1 protein expression in NSCLC cells. Numerical numbers denoted the ratio of integrated optical density (IOD)/IOD of β-actin. B Blocking AS-tDR-007333 suppressed HSPB1 protein expression. C Up-regulation of AS-tDR-007333 promoted HSPB1 gene expression in NSCLC cells. D Knockdown of AS-tDR-007333 decreased HSPB1 gene expression in NSCLC cells. E si-HSPB1 inhibited both HSPB1 and MED29 protein expressions. F Co-IP assay indicated that HSPB1 protein interacted with MED29 protein in A549 cells. G Inhibition of HSPB1 suppressed MED29 gene expression in PC9 and A549 cells. H Rescue assays showed an interaction between AS-tDR-007333 and si-HSPB1 for regulating MED29 promoter activity. I, J Co-transfection of MED29 and si-HSPB1 into NSCLC cells rescued the promoting effect of MED29 on cell proliferation. K Western blot analysis revealed that knockdown of HSPB1 suppressed the expression of H3K4me1 and H3K27ac in NSCLC cells. L ChIP-qPCR results of H3K4me1 signal on different regions of MED29 promoter in the presence or absence of si-HSPB1. M ChIP-qPCR results of H3K27ac expression levels on different regions of MED29 promoter in the presence or absence of si-HSPB1. *P < 0.05; **P < 0.01; ***P < 0.001