Fig 4. Inverse agonist of ERRγ inhibits ACEA-mediated fibrinogen gene expression in Huh7 cells.

(A) GSK5182 decreased ACEA-mediated FGG promoter activity. Huh7 cells were transfected with vectors expression hFGG-luc, and then treated with ACEA (10 μM) and/or GSK5182 (10 μM). (B-E) GSK5182 inhibited ACEA-mediated fibrinogen expression and secretion in human hepatoma cell line. Huh7 and HepG2 cells were treated with ACEA (10 μM) for 12 h. The cell culture medium was replaced, and GSK5182 (10 μM) was added for the final 24 h. Total mRNA and protein were extracted for qPCR (B-C) and western blot analyses (D). Cell culture media were collected to determine fibrinogen levels in Huh7 cells (E). (F) GSK5182 specifically inhibits ERRγ transcriptional activity. Huh7 cells were infected with Ad-GFP, Ad-ERRγ, or Ad-ERRγ Y326A and then treated with GSK5182 (10 μM) for 24 h. Fibrinogen mRNA levels were analyzed by qPCR. * p<0.05, ** p<0.01. All data are representative of at least three independent experiments. Western blot images were cropped with a black cropping line. All gels for Western blot analysis were run under the same experimental conditions. Full uncropped blots are available as S2 Fig. Error bars show SEM.