Af 379 na

CD4, IL-17, IL-4, pSTAT3 (y705), CXCR3, and procollagen levels were analyzed by confocal microscopy. Paraffin-embedded sections were incubated with 10% normal goat serum for 30 min and stained with anti-CD4 (AF-379-NA; R&D Systems), anti-IL-17 (MAB3171-100; R&D Systems), anti-IL-4 (PA5-25165; Thermo Fisher Scientific), anti-pSTAT3 (y705) (ab76315; Abcam), anti-CXCR3 (SC-133087; Santa Cruz Biotechnology, Dallas, TX), and anti-procollagen (ab64409; Abcam) antibodies. Next, the samples were reacted with anti-goat IgG-FITC (SC-2024; Santa Cruz Biotechnology), anti-mouse-IgG1-Alexa Fluor 555 (A21127; Invitrogen, Waltham, MA), anti-rabbit-IgG-PE (4050-09; Southern Biotech, Birmingham, AL), anti-rabbit-IgG APC (A-10931; Invitrogen), anti-mouse-IgG-Alexa Fluor 647 (1031-31; Southern Biotech), and anti-rat IgG-Alexa Fluor 488 (A-11006; Invitrogen)-conjugated secondary antibodies. Nuclei were stained with DAPI (D3571; Thermo Fisher Scientific). The sections were visualized using a confocal microscope (LSM 700; Carl Zeiss, Oberkochen, Germany); the colocalization area and intensity were analyzed using ZEN 2009 software (Carl Zeiss).

The liver tissues were sectioned at 4 µm for histopathology staining, as previously described (17 (link)). For immunohistochemistry (IHC), paraffin sections were incubated with antibodies against IgM (Abcam, #ab134159), IgG (Abcam, #ab109489) following previous work (16 (link)). For immunofluorescent (IF), paraffin sections were incubated with antibodies against rabbit anti-human CXCR5 (Abcam, #ab133706) and goat anti-human CD4 (R&D systems, #AF-379-NA), followed by Alexa Fluor 488 or 555 conjugated secondary antibodies (Invitrogen, Carlsbad, CA). The images were digitized using a Zeiss fluorescence microscope (LSM 710).