Figure 3.
Effect of angiotensin II (A-II), forskolin, and KCNJ5 mutation in HAC15 cells, and DNA methylation on mRNA expression of CLGN in human adrenocortical tumors. Confluent HAC15 cells were serum deprived for 24 h, then incubated with A-II (A), forskolin (B), or no stimulant for 3 h. After aspiration of the media, the cells were harvested, and CLGN and GAPDH mRNA levels were measured (n = 3). (C) Serum-deprived HAC15 cells with or without a KCNJ5 mutation were harvested, and CLGN and GAPDH mRNA levels were measured (n = 3). (D) Serum-deprived HAC15 cells with or without a ATP1A1 mutation were harvested, and CLGN and GAPDH mRNA levels were measured (n = 3). (E) Putative methylation sites numbered from the transcription start site of CLGN gene are shown. (F) A heat map of CLGN methylation levels between NFA (n = 12) and APA (n = 35). There were no differences of CLGN methylation levels between NFA and APA.