fig5

Figure 5. Glucocorticoids/IL-6 acts as the co-activator of Reg3b and Reg3g transcription.
(A) The mRNA levels of Reg3b and Reg3g measured in primary hepatocytes isolated from WT mice and treated with TGF-β, Activin a, or BMP4 for 12 h (n = 5 per group); (B) The mRNA level of Reg3b and Reg3g measured in primary hepatocytes isolated from control and LKO mice and differentially treated with Dex, Hyd, IL6 (n = 4 per group); (C) Volcano plot representation of RNA sequencing data of genes upregulated (red) or downregulated (blue) in the liver from WT mice (non-infected) and WT mice infected with E. coli (12 h postinfection). Gray dots indicate genes that were not significantly upregulated or downregulated; (D) Volcano plot representation of RNA sequencing data of genes upregulated (red) or downregulated (blue) in the liver from E. coli infected WT mice and WT + RU486 mice (12 h post infection). Gray dots indicate genes that were not significantly upregulated or downregulated; (E) The mRNA level of Reg3b and Reg3g in the liver of WT mice and WT + RU486 mice (at non-infected conditions and upon E. coli infection) (n = 6-9 per group); (F) Representative flow cytometry dot plot of Kupffer cells, macrophage and Monocytes in the liver of WT mice and WT + RU486 mice at non-infected conditions and upon E. coli infection (n = 6-9 per group); (G) The corresponding cell proportion/number of Kupffer cells, macrophage and Monocytes in the liver of WT mice and WT + RU486 mice at non-infected conditions and upon E. coli infection (n = 6-9 per group); (H) Representative flow cytometry dot plot of inflammatory monocytes in the blood of WT mice and WT + RU486 mice (at non-infected conditions and upon E. coli infection) (n = 6-9 per group); (I) The corresponding cell proportion/number of inflammatory monocytes in the blood of WT mice and WT + RU486 mice (at non-infected conditions and upon E. coli infection) (n = 6-9 per group); (J) The bacterial CFU counts in the peritoneal fluid, blood and liver of WT and WT + RU486 mice 12 h post E. coli infection (n = 6-9 per group); (K) Serum ALT and AST levels of WT mice and WT + RU486 mice at non-infected conditions and upon E. coli infection (n = 6-9 per group). Circles correspond to individual mouse. Results are represented as mean ± SD. The P values in (A) were determined using one-way ANOVA. The P values in (B, E, G, H and I) were determined using two-way ANOVA. The P values in (J) were determined using unpaired Student’s t-tests. ^*P < 0.05; ^**P < 0.01; ^***P < 0.001. NS: Not significant; PBS: phosphate buffered saline; KC: Kupffer cell; DMSO: dimethyl sulfoxide; LKO: liver-specific Smad4 knockout; ALT: alanine transaminase; AST: aspartate transaminase; WT: wild type; TGF-β: transforming growth factor-β; BMP4: bone morphogenetic protein 4; Dex: dexamethasone; Hyd: hydrocortisone; IL-6: interleukin-6; E. coli: Escherichia coli; RU486: mifepristone; CFU: colony forming unit.