PROK1 increases CTGF mRNA and protein expression, via Gq, PLC, cSrc, EGFR and MEK signalling in PROKR1-Ishikawa cells. (A) CTGF mRNA and (B) protein expression in PROKR1-Ishikawa cells after treatment with 40 nM PROK1. CTGF mRNA expression peaked after 12 h and protein expression peaked after 48 h of PROK1 treatment. (C) Expression of CTGF mRNA was measured after treatment with vehicle or 40 nM PROK1 in the absence or presence of the specific cell signalling inhibitors YM-254890 (Gq inhibitor; YM), U73122 (PLC inhibitor; U7), PP2 (cSrc inhibitor), GM1489 (matrix metalloproteinase inhibitor; GM), AG1478 (EGFR inhibitor; AG), PD98059 (MEK inhibitor; PD) or inhibitor of INCA6 (NFAT cells, inhibitor, INCA 6). PROK1-induced CTGF mRNA expression was significantly inhibited in the presence of these inhibitors with the exception of INCA 6. Each bar represents the mean ± SEM of at least three independent experiments. ***P < 0.001, **P < 0.01, *P < 0.05 compared with vehicle treated cells (A and B) or PROK1 treatment alone (C), as determined by one-way ANOVA.