Fig. 3

Barx homeobox 1 (BARX1) regulates HSPA6 expression directly in OS cells. A RNAs derived from MG63 cells transfected with pcDNA-BARX1 or vectors were extracted and subjected to RNA sequencing. Heatmap of the DEGs in MG63 cells with BARX1 knockdown compared with controls. B MG63 cells were transfected with either the overexpression plasmid of BARX1 or the negative control (NC). After 48 h, RNA was extracted, and qPCR was performed to detect the expression of DNAJB1, HSPAB1, HSPA6, SERPINH1, CLDN6, and ARC. C The binding sites of BARX1 on the HSPA6 promoter. A dual-luciferase assay was used to analyse the effect of BARX1 on the HSPA6 promoter. D Mutation of the predicted binding sites of BARX1 on the HSPA6 promoter. E A dual-luciferase assay was used to analyse the effect of BARX1 on the HSPA6 promoter with different mutational sites. F HOS and MG63 cells were transfected with the BARX1 overexpression plasmid for 48 h. Western blotting was performed to determine the HSPA6 protein level