Fig. 3: CypA is a proviral factor in the presence of TRIM5.
From: TRIM5α restricts poxviruses and is antagonized by CypA and the viral protein C6
a, Plaque image 24 hpi of T-REx-293 (WT) and derivative CypA−/− (KO1 and KO2) cells (top), and T-REx-293 TRIM5−/− and derivative TRIM5−/−CypA−/− (double knockout 1 (DKO1) or DKO2) cells (bottom) with VACV A5–GFP. b, Plaque area quantification of cells described in the top panels of a. n ≥ 107 per condition. c, Infectious VACV titres following infection of T-REx-293 (WT) and derivative CypA−/− (KO1 and KO2) cells at 5 PFU per cell for 16 h. n = 3 per condition. d, Plaque area quantification of cells described in the bottom panels of a. n ≥ 236 per condition. e, Infectious VACV titres following infection of T-REx-293 (WT) and derivative TRIM5−/−, and TRIM5−/−CypA−/− (DKO1 and DKO2) cells at 5 PFU per cell for 16 h. n = 3 per condition. f, Plaque area quantification 2 days post-infection (dpi) of HeLa and derivative TRIM5−/− cells with VACV A5–GFP with the indicated concentrations of CsA. n ≥ 13 per condition. g, Plaque area quantification 24 hpi of T-REx-293 CypA−/− cells engineered to inducibly express (+Dox) TAP-tagged WT CypA and mutants R55A and F113A with VACV A5–GFP. n ≥ 140 per condition. h, Infectious VACV titres following infection of cells described in g at 5 PFU per cell for 16 h. n = 3 per condition. In g,h, cells were treated with 150 ng ml−1 doxycycline from seeding for 24 h to express tagged proteins before harvest or infection. Data shown in a,b,d,f,g are representative of three independent experiments, and in c,e,h are from two independent experiments. Data from b–f were analysed using one-way Welch’s ANOVA test (P < 0.0001 in f), and pairwise comparisons in f were performed using post-hoc Dunnett’s T3 multiple comparisons test. Data from g,h were analysed using two-tailed unpaired Student’s t-test. Analyses were performed on GraphPad Prism. Data are mean ± s.e.m. (b–h).
