Fig. 5.

IFIT1 mediated PD-L1 stabilization via regulating PD-L1 ubiquitination. A. Co-immunoprecipitation (Co-IP) assay detecting IFIT1 in RKO cell lysates immunoprecipitated with anti-PD-L1. B. RKO cells were overexpressed with FLAG_IFIT1 and HA_PD-L1. Co-IP assay detecting exogenous IFIT1 (FLAG) in RKO cell lysates immunoprecipitated with anti-HA. C-D. RKO cells transfected with control siRNAs and IFIT1 siRNAs were treated with cycloheximide (CHX) (50 µg/mL) at the indicated time points. Immunoblot of PD-L1 at indicated time points. E. Quantification of relative remaining PD-L1 in C-D. F. Immunoblot of PD-L1 in RKO cells transfected with IFIT1 siRNAs and treated with MG132 (10uM, 6 hours). G. Immunoblot of PD-L1 in RKO cells transfected with IFIT1 siRNAs and treated with CQ (25uM, 6 hours). H. Immunoblot of PD-L1 in RKO cells transfected with IFIT1 siRNAs and treated with 3-MA (6 hours). I-J. Immunoblot of cell lysates immunoprecipitated with anti-PD-L1 in RKO cells transfected with the indicated constructs.