FIGURE 6.

Knockdown of USP17 decreases RORγt protein and Th17-related gene expression in Th17 cells. A, Myc-tagged USP17 and shUSP17-1, ShUSP17-2, and ShUSP17-3 were cotransfected into 293T cells. Protein levels were detected by immunoblotting (IB) with anti-Myc antibodies. B and C, naïve CD4⁺T cells were polarized under Th17 conditions. After polarization, Th17 cells and FLAG-RORγt-Jurkat cells were transduced with a lentivirus containing shCK (control) or shUSP17-3. Western blotting was performed after screening for the cells with G418 or puromycin. Quantification of RORγt or USP17 protein levels was done with ImageJ software (normalized to GAPDH intensity). D, RT-PCR was performed after selecting for screening for the cells with G418 or puromycin. Data are representative of more than three independent experiments. Error bars show mean ± S.D.