Fig. 2: TRIM5α is a VACV restriction factor.
From: TRIM5α restricts poxviruses and is antagonized by CypA and the viral protein C6
a, Infectious VACV titres following infection of T-REx-293 (WT) and derivative TRIM5−/− (KO1 or KO2) cells with VACV-WR at 5 plaque-forming units (PFU) per cell for 16 h. n = 3 per condition. b, Plaque image (left) and plaque area quantification (right) 24 hpi for the cells described in a with VACV A5–GFP. n ≥ 114 per condition. c, Plaque area quantification 24 hpi with VACV A5–GFP of T-REx-293 TRIM5−/− cells engineered to inducibly express (+Dox) TAP-tagged TRIM5α, TRIM5γ and TRIM5δ isoforms. n ≥ 87 per condition. EV, empty vector. d, Infectious VACV titres following infection of cells described in c at 5 PFU per cell for 16 h. n = 3 per condition. e, Plaque area quantification 24 hpi with VACV A5–GFP of T-REx-293 TRIM5−/− cells engineered to inducibly express (+Dox) the following TAP-tagged TRIM5α mutants: L19R, N70A, R119E and ΔSPRY. n ≥ 54 per condition. f, Infectious VACV titres following infection of cells described in e at 5 PFU per cell for 16 h. n = 3 per condition. In c,d, cells were treated with 150 ng ml−1 doxycycline from seeding for 24 h to express tagged proteins before harvest or infection. In e,f, doxycycline was added at 24 h after seeding. Data shown in b,c,e are representative of three independent experiments, and in a,d,f are from two independent experiments. Data from a,b were analysed using one-way Welch’s analysis of variance (ANOVA) test. Data from c–f were analysed using two-tailed unpaired Student’s t-test. Analyses were performed on GraphPad Prism. Data are mean ± s.e.m.
