Fig. 6: Signal pathways and TFs involved in IL6-induced hepatocyte expansion.

a Western blot analysis of STAT3, p-STAT3, AKT, p-AKT, ERK1/2, p-ERK1/2 in hepatocytes treated with IL6, EGF/HGF or the combination for 2, 4 and 6 h. GAPDH was used as loading control. b Cell numbers of hepatocytes cultured in IL6-HCM and a number of pathway modulators, including Crytotanshinone and BP-1-102 (JAK/STAT3 inhibitor), MK2206 and GSK2141795 (PI3K/AKT inhibitor), PD0325901 (MARK/ERK1/2 inhibitor), and TGFβ1 for 14 days. Data are Means ± SEM (n = 4 independent experiments). ^###p < 0.001 vs. first bar, *p < 0.05, **p < 0.01, ***p < 0.001 vs. second bar (two-tailed unpaired Student’s t test). c Quantitative RT-PCR analysis of Barx2, Elf3, Mxd3 and FoxM1 in hepatocytes cultured in IL6-HCM for 14 days, and IL6-iHPCs (P10). Data are Means ± SEM (n = 3 independent experiments). d–g Quantitative RT-PCR analysis (left) of Barx2, Elf3, Mxd3 and FoxM1 expression and cell number (right) of IL6-iHPCs (P10) 5 days after transfection of shBarx2, shElf3, shMxd3, shFoxM1 or scramble sequence. Data are Means ± SEM (n = 3 independent experiments). **p < 0.01, ***p < 0.001 (two-tailed unpaired Student’s t test). Representative images (h) and cell number (i) of IL6-iHPCs (P10) with combination knockdown of Barx2, Elf3, Mxd3 and FoxM1 in IL6-HCM. Data are Means ± SEM (n = 3 independent experiments). **p < 0.01, ***p < 0.001 (two-tailed unpaired Student’s t test). j Quantitative RT-PCR analysis of Barx2, Elf3, Mxd3 and FoxM1 expression in IL6-iHPCs (P10) cultured in IL6-HCM with various pathway inhibitors (Cry, BP, MK, GSK and PD) for 5 days. Data are Means ± SEM (n = 3 independent experiments). ^##p < 0.01, ^###p < 0.001 vs. first bar, *p < 0.05, **p < 0.01, ***p < 0.001 vs. second bar (two-tailed unpaired Student’s t test). Scale bars represent 100 µm. Source data and exact p values are provided in a Source data file.