Figure 5.

HOXA5 regulates WNK1 expression via direct binding to its promoter. (A, C) The mRNA level of HOXA5 or WNK1 in transfected macrophages was detected by qRT-PCR. n = 3. (B, D) The protein level of HOXA5 or WNK1 in transfected macrophages was detected by Western blotting with quantitative analysis. n = 3. (E) The putative binding site between HOXA5 and WNK1 promoter. n = 3. (F) The direct interaction between HOXA5 and WNK1 promoter in macrophages was detected by ChIP assay. n = 3. PCR products were assessed by agarose gel electrophoresis. Anti-H3 antibody and normal IgG served as a positive and negative control, respectively. n = 3. (G) The relative luciferase activity in transfected macrophages was assessed by dual luciferase assay. n = 3. (H&I) The mRNA and protein levels of WNK1 in transfected macrophages were detected by qRT-PCR and Western blotting with quantitative analysis, respectively. n = 3. *, P < 0.05; **, P < 0.01; ***, P < 0.001 as assessed by Student's t-test (A&B) or One-way ANOVA with Turkey post hoc test (C–I).