Effect of mineralocorticoid receptor (MR) and aldosterone in HEK293T cells expressing Kcnn4 genomic clone.

Regions of the Kcnn4 genes showing positive response in a ChIP assay and indicated by bars in Figure-1C (clone-1, clone-3, clone-5, clone-6, clone-7 and clone-8) were sub-cloned in pGL4.23 and tested for the presence of MR/aldosterone (mineralocorticoid)-responsive enhancers (MRE). The HEK293T cells were transfected with pGL4.23 or pGL4.23 containing Kcnn4 genomic clones. The cells transfected with Kcnn4 genomic clones were also transfected with MR. Luciferase activity was measured in the cells transfected with the Kcnn4 genomic clones with (hatched bars) and without (grey bars) MR transfection. The luciferase activity in the cells transfected with the Kcnn4 genomic clones and MR was also measured in the presence of 10 nM aldosterone (black bars). For clarity, luciferase activity measured in the presence of a plasmid, MR and aldosterone are shown in cells transfected with pGL-4.23, clones-3, -5 and -6. Results presented represent mean ± SE from 3 different experiments. *p<0.05– compared to pGL4.23.