Goat anti cxcl9 antibody

Immunofluorescence was performed as previously described52 (link). Mouse kidneys were fixed by perfusion through the left ventricle with 0.2 M periodate lysine and 2% paraformaldehyde in PBS. Tissue samples were soaked for several hours in 20% sucrose in PBS, embedded in Tissue-Tek OCT Compound (Sakura Finetechnical), and snap-frozen in liquid nitrogen. Goat anti-CXCL9 antibody was purchased from R&D Systems. Fluorescent lotus tetragonolobus lectin (LTL) was purchased from Vector Laboratories. Alexa fluor (Molecular Probes; Invitrogen) was used for secondary antibodies. Immunofluorescent images were obtained using the Leica TCS SP8 laser-scanning confocal microscope system.

Immunoblotting was performed as previously described53 (link). For immunoblotting, we used whole lysates of entire kidney samples without the nuclear fraction (600 g) and crude lysates of HK2 cell samples (15,000 g). Goat anti-CXCL9 antibody was purchased from R&D Systems. Rabbit anti-STAT1 antibody, rabbit anti-phosphorylated STAT1 (Tyr⁷⁰¹) antibody, rabbit anti-JAK1 antibody, rabbit anti-phosphorylated JAK1 (Tyr^1022/1023) antibody, rabbit anti-JAK2 antibody, rabbit anti-phosphorylated JAK2 (Tyr^1007/1008) antibody, and rabbit anti-Histone H3 antibody were purchased from Cell Signaling. Rabbit anti-IFNGR1 antibody was purchased from Santa Cruz Biotechnology. Rabbit anti-β-ACTIN antibody was purchased from Sigma-Aldrich. Alkaline phosphatase-conjugated anti-IgG antibody (Promega) was used as the secondary antibody, and Western Blue (Promega) was used to detect the signals. The band intensities of the western blots were quantified using Image J software (NIH).