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From: Hepatic Smad4 mediates antibacterial defense via macrophage recruitment

Figure 3. The supplementation of Reg3b and Reg3g exert protective effects on LKO mice during infection.
(A) The LKO mice were injected intraperitoneally with 1ug of mouse recombinant Reg3b and Reg3g (rReg3b and rReg3g) or PBS at the time of E. coli infection. The mice were sacrificed 12 h post infection and analysis for bacterial CFU counts in the peritoneal fluid, blood and liver (n = 5 per group); (B) Fluorescence microscopy of liver tissues of control, LKO + PBS, LKO + rReg3b and Reg3g mice infected with GFP-labeled E. coli; (C) Serum ALT and AST levels of control, LKO + PBS, LKO + rReg3b and rReg3g mice were measured 12 h post infection (n = 4 per group); (D) Representative hematoxylin and eosin staining of liver tissue in control, LKO + PBS and LKO + rReg3b and rReg3g mice 12 h post infection; (E) Schematic diagram illustrating the experimental design and potential anti-bacteria mechanism of rReg3b and rReg3g treatment for LKO mice; (F) The CFU counts of E. coli after exposure to recombinant Reg3b and Reg3g in vitro (n = 3 per group); (G) Bacterial killing capacity of control, LKO macrophages and LKO macrophages treated with rReg3b and rReg3g, following stimulation with E. coli (n = 5 per group); (H) Representative flow cytometry dot plot of macrophages, monocytes and myeloid cells in the peritoneal fluid of control (n = 4) and LKO mice (n = 5); (I) Representative cell proportion and number of macrophages, monocytes and myeloid cells in the peritoneal fluid of control (n = 4) and LKO mice (n = 5). Circles correspond to individual mouse. Results are represented as mean ± SD. The P values in (A and C) were determined using one-way ANOVA. The P values in (F) were determined using unpaired Student’s t-tests. The P values in (G and I) were determined using two-way ANOVA. ^*P < 0.05; ^**P < 0.01; ^***P < 0.001. NS: Not significant; CFU: colony forming unit; PBS: phosphate buffered saline; LKO: liver-specific Smad4 knockout; GFP: green fluorescent protein; E. coli: Escherichia coli; ALT: alanine transaminase; AST: aspartate transaminase.