Fig 2: BRG1-dependent hepatocyte-derived CCL7 promotes macrophage migration. (A) HepG2 cells were transfected with siRNA targeting BRG1 or scrambled siRNA (SCR) followed by treatment with LPS for 6 h. Conditioned media were collected and chemotaxis assay was performed as described in Methods. (B) HepG2 cells were transfected with siRNA targeting BRG1 or scrambled siRNA (SCR) followed by treatment with palmitate for 12 h. Conditioned media were collected and chemotaxis assay was performed as described in Methods. (C) Primary hepatocytes were isolated from WT or LKO mice and treated with or without LPS for 6 h. Conditioned media were collected and chemotaxis assay was performed as described in Methods. (D) Primary hepatocytes were isolated from WT or LKO mice and treated with palmitate for 12 h. Conditioned media were collected and chemotaxis assay was performed as described in Methods. (E) HepG2 cells were infected with BRG1 adenovirus (Ad-BRG1) or the control adenovirus (Ad-GFP) followed by treatment with LPS for 6 h. Conditioned media were collected and chemotaxis assay was performed in the presence or absence of a CCL7-neutralizing antibody. (F) HepG2 cells were infected with BRG1 adenovirus (Ad-BRG1) or the control adenovirus (Ad-GFP) followed by treatment with palmitate for 12 h. Conditioned media were collected and chemotaxis assay was performed in the presence or absence of a CCL7-neutralizing antibody. (G) Primary hepatocytes were infected with BRG1 adenovirus (Ad-BRG1) or the control adenovirus (Ad-GFP) followed by treatment with LPS for 6 h. Conditioned media were collected and chemotaxis assay was performed in the presence or absence of a CCL7-neutralizing antibody. (H) Primary hepatocytes were infected with BRG1 adenovirus (Ad-BRG1) or the control adenovirus (Ad-GFP) followed by treatment with palmitate for 12 h. Conditioned media were collected and chemotaxis assay was performed in the presence or absence of a CCL7-neutralizing antibody.

Fig 3: BRG1 interacts with AP-1 to activate CCL7 transcription in a redox-sensitive manner. (A) CCL7 promoter-luciferase constructs were transfected into HepG2 cells with or without BRG1 followed by treatment with LPS or PA. Luciferase activities were normalized by protein concentration and GFP fluorescence. (B) HepG2 cells were transfected with siRNA targeting AP-1 or scrambled siRNA (SCR) followed by treatment with LPS or PA. ChIP assays were performed with anti-BRG1 or IgG. (C) HepG2 cells were treated with LPS or PA in the presence or absence of NAC. ChIP assays were performed with anti-c-Jun, anti-c-Fos, or anti-BRG1. (D) HepG2 cells were treated with LPS or PA in the presence or absence of NAC. Immunoprecipitation was performed with anti-BRG1 or IgG. (E) C57/BL6 mice were injected with LPS in the presence or absence of NAC for 12 h. Immunoprecipitation was performed with anti-BRG1. (F) C57/BL6 mice were fed an MCD diet in the presence or absence of NAC for 4wk. Immunoprecipitation was performed with anti-BRG1. (G) HepG2 cells were treated with LPS or PA for 12 h. Re-ChIP assay was performed with indicated antibodies.

Fig 4: CK2-mediated BRG1 phosphorylation promotes its interaction with AP-1. (A, B) C57/BL6 mice were injected with LPS in the presence or absence of NAC for 12 h. CK2 expression levels in the liver were examined by qPCR and Western. Immunoprecipitation was performed with anti-BRG1. (C, D) C57/BL6 mice were fed an MCD diet in the presence or absence of NAC for 4wk. CK2 expression levels in the liver were examined by qPCR and Western. Immunoprecipitation was performed with anti-BRG1. (E) HepG2 cells were treated with LPS or PA in the presence or absence of NAC. CK2 expression levels were examined by qPCR. (F) Primary murine hepatocytes were treated with LPS or PA in the presence or absence of NAC. CK2 expression levels were examined by qPCR. (G) HepG2 cells were transfected with siRNA targeting CK2 or scrambled siRNA (SCR) followed by treatment with LPS or PA. Immunoprecipitation was performed with anti-BRG1. (H) HepG2 cells were transfected with siRNA targeting CK2 or scrambled siRNA (SCR) followed by treatment with LPS or PA. ChIP assay was performed with anti-BRG1. (I) HepG2 cells were transfected with siRNA targeting CK2 or scrambled siRNA (SCR) followed by treatment with LPS or PA. Re-ChIP assay was performed with indicated antibodies. (J-L) HepG2 cells were transfected with siRNA targeting CK2 or scrambled siRNA (SCR) followed by treatment with LPS or PA. CCL7 expression was examined by qPCR and ELISA. Conditioned media were collected and chemotaxis assay was performed as described in Methods.

Fig 5: BRG1-mediated CCL7 induction and macrophage infiltration is redox-sensitive. (A-C) BRG1 conditional knock-in (LKI) mice and wild type littermates were injected with LPS for 12 h with or without NAC. CCL7 expression was examined by qPCR and ELISA. Macrophages were stained with anti-CD68. (D-F) BRG1 conditional knock-in (LKI) mice and wild type littermates were fed an MCD diet for 4 weeks with or without NAC. CCL7 expression was examined by qPCR and ELISA. Macrophages were stained with anti-CD68. (G-I) Primary hepatocytes were infected with BRG1 adenovirus (Ad-BRG1) or the control adenovirus (Ad-GFP) followed by treatment with LPS and/or NAC. CCL7 expression was examined by qPCR and ELISA. Conditioned media were collected and chemotaxis assay was performed as described in Methods. (J-L) Primary hepatocytes were infected with BRG1 adenovirus (Ad-BRG1) or the control adenovirus (Ad-GFP) followed by treatment with palmitate and/or NAC. CCL7 expression was examined by qPCR and ELISA. Conditioned media were collected and chemotaxis assay was performed as described in Methods. Scale bar, 100 µm.