Anti actinin 4

The protein lysates from the cells and kidney tissues were prepared following standard protocols, and the protein content was determined using the BCA protein assay kit (ThermoFisher). And then the proteins samples were separated by Bis-Tris Gel (Invitrogen) and transferred onto PVDF membranes (Millipore) using a wet- transfer system. Membranes were blocked in 5% BSA in TBS-T for 1 h at room temperature and were incubated with primary antibodies overnight at 4 °C. Then membranes were washed and incubated with secondary horseradish peroxidase-conjugated antibodies for 2 h at room temperature, and the signals were detected using an ECL advanced system (GE Healthcare). Intensity values expressed as the relative protein expression were normalized to β-actin. Primary antibodies used were anti-CD68 (ab955, Abcam); anti-F4/80 (ab6640, Abcam); anti-CD206 (ab64693, Abcam); anti-actinin 4 (15145, Cell Signaling Technology); anti-integrin α4 (8440, Cell Signaling Technology); anti-integrin β1 (4706, Cell Signaling Technology); anti-integrin αL (ab186873, Abcam); anti-integrin β2 (ab185723, Abcam); anti-NF-κB p65 (8242, Cell Signaling Technology); anti-NF-κB p-p65 (3033, Cell Signaling Technology); anti-GR (12041, Cell Signaling Technology); anti-β-actin (ab8226, Abcam). Secondary HRP-conjugated antibodies used were anti-mouse IgG, anti-rabbit IgG, and anti-rat IgG (Abcam).

The primary antibodies used in this study were as follows: anti-CD63 (dilution 1:1000, Santa Cruz Biotechnology, Santa Cruz, CA, USA), anti-CD81 (dilution 1:1000, MBL, Nagoya, Japan), anti-TSG101 (dilution 1:1000, Abcam, Cambridge, MA), anti-flotillin-1 (dilution 1:1000, Cell Signaling Technology, Danvers, MA), anti-actinin-4 (dilution 1:1000, Cell Signaling Technology), anti-β-actin (dilution 1:5000, Sigma-Aldrich), anti-CLDN11 (dilution 1:1000, R&D Systems, Minneapolis, MN), anti-vimentin (dilution 1:1000 in western blotting and 1:100 in immunocytochemistry, Cell Signaling Technology), anti-ZO-1 (dilution 1:1000, Cell Signaling Technology), and anti-Snail (dilution 1:1000, Cell Signaling Technology). The secondary antibodies used in this study were as follows: horseradish peroxidase (HRP)-conjugated horse anti-mouse IgG (dilution 1:1000, Cell Signaling Technology), HRP-conjugated goat anti-rabbit IgG (dilution 1:1000, Cell Signaling Technology), and Alexa Fluor 488-conjugated goat anti-rabbit IgG (dilution 1:1000, Thermo Fisher Scientific).