Fig. 3: Smad4 deficiency in hepatocytes attenuated liver inflammation and CXCL1 secretion.
Smad4fl/fl and AlbSmad4−/− mice were fed an HFD for 4 months to establish the NAFLD model (n = 5 per group). Data are representative of at least three independent experiments. A Immunofluorescence detection and statistical analysis of F4/80, CD11b, and Gr1 expression in NAFLD liver tissues. Scale bar, 50 µm **P < 0.01; ***P < 0.001. B Double immunofluorescence staining for CXCL1 (red) and Albumin (green) in liver tissue. Scale bar, 50 μm ***P < 0.001. C The relative mRNA expression of CXCL1 in liver tissues was measured using RT-qPCR, *P < 0.05. D–I AML12 cells were transfected with si-NC or si-Smad4, respectively, cultured with BSA or palmitic acid (PA, 500 μM) for 24 h. D The protein levels of Smad4 in control si-NC and si-Smad4 AML12 cells were determined using Western blot. E Relative mRNA expression of CXCL1 was measured using RT-qPCR in AML12 cells *P < 0.05. F ELISA verification of culture supernatants from AML12 cells *P < 0.05. G–I AML12 cells transfected with lentiviral vectors for control or Smad4 knockdown were then cultured with BSA or PA (500 μM) for 24 h. G Western blot analysis of Smad4 protein expression in sh-GFP and sh-Smad4 AML12 cells. H The mRNA levels of CXCL1 were measured by RT-qPCR in AML12 cells ***P < 0.001. I Secretory protein levels of CXCL1 in AML12 cell medium were determined using ELISA. **P < 0.01.
