Fig. 1
From: HIV-1 protease cleaves the serine-threonine kinases RIPK1 and RIPK2
HIV PR binds RIP kinase family members. a HEK293T cells (1 × 106) were transiently transfected with indicated expression plasmids (1 μg each). Various Strep-FLAG- (SF-) tagged HIV-1 proteins were expressed along with Myc-tagged RIPK2. After 24 h, proteins were affinity purified from cleared cell lysates with Dynabeads Streptavidin. Affinity purifications (APs) or total lysates were subjected to SDS-PAGE and Western blot (WB) analysis. Proteins were revealed using the indicated antibodies. b HIV PR binds to endogenous RIPK2. Catalytically inactive (D25N) SF-tagged HIV PR was expressed in HEK293T cells. Mock-transfected cells (-PR) were used as control. Complexes were purified as above. APs or total lysates were subjected to SDS-PAGE and Western blot (WB) analysis. Proteins were revealed using antibodies against RIPK2 (BD Transduction Laboratories) or β-actin (Sigma-Aldrich)