Skip to main content
Fig. 6 | Retrovirology

Fig. 6

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

Fig. 6

Processing by HIV-1 PR disrupts RIPK1 function. a HEK293T cells harboring a stable NF-κB luciferase reporter were transfected by lipofection with an expression plasmid encoding RIPK1, or empty pcDNA. Cells were co-transfected with or without PR (20 ng/well), then cultured with or without SQV. After 24 h, cells were lysed by addition of Steady-GloR (Promega) and luciferase activity was measured using a standard plate luminometer. b Protein levels in a were analyzed using standard SDS-PAGE/Western blotting techniques. Proteins were revealed using antibodies against c-Myc, β-actin, or HIV-1 PR. Results are from a single experiment and are representative of at least three separate experiments. c, d Mammalian 2-hybrid system. c HEK293T cells were transfected with plasmids encoding RIPK1 fused to the GAL4 DNA-binding domain (RIPK1 only), RIPK3 fused to the GAL4 transcription activation domain (RIPK3 only), or a combination thereof (RIPK1/3). Cells were co-transfected with (+) or without (−) PR in the absence (n.t.) or presence (+) of SQV. d HEK293T cells were transfected with plasmids encoding p53 fused to the GAL4 DNA-binding domain (p53 only), large T antigen fused to the GAL4 transcription activation domain (T only), or a combination thereof (p53/T). Values represent mean ± standard deviation of triplicate cultures (mean ± SD, n = 3). Statistical significance was determined by unpaired t-tests, ****P ≤ 0.0001, ***P ≤ 0.001

Back to article page