Fig. 6
From: Cyclophilin A enables specific HIV-1 Tat palmitoylation and accumulation in uninfected cells
Cyclophilin A and FKBP12 are required for Tat palmitoylation and Tat-mediated inhibition of PI(4,5)P2-dependent membrane traffic. a PC12 cells were transfected with Tat-FLAG before treatment for 6 h with 1 µM FK506, 2 µM CSA, 1 µM rapamycin or solvent. Palmitoylation was then assayed using acyl-biotin exchange before band quantification. Results are expressed as biotin/Tat intensity ratio (% of control). b Jurkat T-cells were transfected with Tat before adding drugs for 6 h and assaying Tat secretion by ELISA. c PC12 cells were transfected with GH before drug treatment for 6 h and GH secretion assay. d Human MDMs were treated with the drug for 5 h before assaying phagocytosis of IgG-coated beads. e PC12 cells were transfected with the indicated siRNA before lysis then anti-CypA or anti-FKBP12 then anti-αtubulin western blot. f PC12 cells were cotransfected with the indicated siRNA and Tat-FLAG before detecting Tat palmitoylation using acyl-biotin exchange. (HA, hydroxylamine). g Jurkat cells were cotransfected with Tat and the indicated siRNA (silencing efficiency is shown in Supplementary Fig. 12b) and Tat secretion was assayed after 48 h. Data are mean ± SEM, n = 3 independent experiments. The significance of differences with controls was assessed using ANOVA, one-way (a, b, g) or two-way (c, d) (***p < 0.001; **p < 0.01; *p < 0.05)
