Immunoblotting Analysis of Naïve CD4+ T Cells

Western blotting analyses were performed with whole cell lysates of antagomiR-31 or antgoNC treated naïve CD4+ T cells according to previously published procedures (25 (link)). Following antibodies were used: mouse anti-actin antibody (1:3,000; Cell Signaling Technology, #3700S), rabbit anti-STAT1 Ab (1:1,000; Cell Signaling Technology, #9172S), and rabbit anti-phospho-STAT1 Ab (Cell Signaling Technology, #9167S). The immunoblots was imaged and analyzed using Image Studio (LI-COR Biosciences). The anti-human IFN-γ antibody (clone B27) used for neutralization of IFN-γ in the cell culture medium and the IgG1κ isotype control antibody (Clone MG1-45, Cat. No. 401409) were purchased from BioLegend.