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Fig. 5 | Retrovirology

Fig. 5

From: HIV-1 protease cleaves the serine-threonine kinases RIPK1 and RIPK2

Fig. 5

RIPK1 and RIPK2 are cleaved during HIV-1 infection. a, b HEK293T were transiently transfected with expression plasmids (0.5 μg/well) encoding Myc-tagged RIPK1, RIPK2, or RIPK3, respectively. After 24 h, cells were infected with different MOIs of VSV-G pseudotyped HIV-1 NL4.3 in the absence or presence of SQV (5 μM) or AZT (10 μM). Cell lysates were prepared 8 or 24 h after infection and subjected to SDS-PAGE and immunoblotting (WB). Proteins were revealed using anti-Myc antibody. c, e Endogenous RIPK1 is cleaved during HIV-1 infection. c HEK293T were infected with different MOIs of VSV-G pseudotyped HIV-1 NL4.3. Cell lysates were prepared 24 h after infection and subjected to SDS-PAGE and immunoblotting (WB). Proteins were revealed using antibodies against RIPK1 (rabbit monoclonal antibody from Cell Signaling) or β-actin. d Sup-T1 cells or e primary activated CD4+ T cells were infected with increasing MOIs of replication-competent HIV-1 NL4.3. Cells were cultured with or without SQV (5 μM), Raltegravir (RAL), or Nevirapine (NVP), respectively. Cell lysates were prepared 48 h after infection and subjected to SDS-PAGE and WB. Proteins were revealed using antibodies against RIKP1 (rabbit monoclonal antibody from Cell Signaling), p24, or β-actin

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