Fig. 10: Increased binding between GATA4 and the Gαi3 promoter in bladder cancer tissues and cells.

The priBlCa-1 primary bladder cancer cells were subjected to stable transduction with GATA4-specific lentiviral shRNAs (shGATA4-S1, shGATA4-S2, representing different sequences), a scramble control shRNA (“shC”), a lentiviral GATA4-overexpressing construct (“oeGATA4”) or an empty vector (“Vec”). The expression levels of targeted mRNAs and proteins were subsequently measured (A–D). Chromatin immunoprecipitation (ChIP) assays presented the relative levels of GATA4-bound Gαi3 promoter in primary (priBlCa-1, priBlCa-2, priBlCa-3) and immortalized (T24) bladder cancer cells, or in the bladder epithelial cells (priBEC-1 and priBEC-2) (E) as well as in the designated bladder cancer tumor tissues (“T1/T2/T3”) and matched adjacent normal bladder epithelial tissues (“N1/N2/N3”) (F), with results quantified. Parental control cells were denoted as “Pare”. Data were presented as mean ± standard deviation (SD). n = 5 stands for five biological repeats. Statistical significance was indicated by *P < 0.05 compared to the “shC”, “Vec”, “priBEC-1” or “N” group. Each experiment was repeated five times, yielding consistent results.