Extended Data Fig. 3: CypA is a pro-viral factor in the presence of TRIM5.
From: TRIM5α restricts poxviruses and is antagonized by CypA and the viral protein C6
a) Immunoblot showing CypA levels in T-REx-293 (WT) and derivative CypA-/- (KO1/2) cells. b) Immunoblot showing TRIM5 and CypA levels in T-REx-293 (WT) and derivative TRIM5-/-, and TRIM5-/-CypA-/- (DKO1/2) cells. c) Infectious VACV titres following infection of cells described in a) at 0.01 PFU/cell for 24 h. n = 3/condition. d) Infectious VACV titres following infection of cells described in b) at 0.01 PFU/cell for 24 h. n = 3/condition. e) Plaque image of Fig. 3f. Plaques were selected at random, imaged using AxioVision 4.8 and measured using ImageJ. f) Immunoblot showing inducible expression (+Dox) of TAP-tagged WT CypA and mutants R55A and F113A, introduced into T-REx-293 CypA-/- cells. g) Plaque image 24 hpi of cells indicated in f) with VACV A5-GFP. h) Infectious VACV titres following infection in cells described in f) at 0.01 PFU/cell for 24 h. n = 8/condition for EV, n = 4/condition for CypA WT and n = 6/condition for R55A and F113A. Data shown in e) and g) are representative of three independent experiments, and c), d) and h) are from two. Data from c) and d) were analysed using One-Way Welch’s ANOVA test. Data from h) was analysed using two-tailed unpaired Student’s t-test. Analyses were performed on GraphPad Prism. Mean ± s.e.m.
