Fig. 7. NFATc2 activates the expression of NOTCH3 and ABCG2 by binding to their promoters.

A Schematic depiction of the NOTCH3 and ABCG2 promoters that are fused to a luciferase reporter gene. B ChIP assay was performed using control IgG and anti-NFATc2 antibody in A549 cells. The precipitated DNA was amplified with primer pairs flanking the putative NFATc2-binding regions of indicated promoters as depicted in (A) by PCR. C, D Quantitative analysis of NFATc2-binding to the indicated promoter regions of NOTCH3 and ABCG2 was determined by ChIP assay followed by qPCR. E, F The luciferase activities of NOTCH3 and ABCG2 reporters in the indicated cell fractions. The indicated reporters with wild type or mutant NFATc2-binding consensus sequences were co-transfected with PRL-TK plasmid, which serves as an internal control, into sorted α2δ1⁻ and α2δ1⁺ A549 cells. Luciferase values were normalized to Renilla reporter activities. G, H The luciferase activities of wild type NOTCH3 and ABCG2 reporters in the indicated cell fractions treated with control vehicle DMSO or CsA (1 µmol/L). All data are presented as mean ± SD of three independent experiments (n = 3). *Student’s t-test.