Fig. 1
From: Primate TRIM34 is a broadly-acting, TRIM5-dependent lentiviral restriction factor
Diverse primate TRIM34 orthologues restrict SIVAGM-SAB, SIVAGM-SAB, and SIVMAC capsids in the presence of TRIM5α. a. THP-1 TRIM34 clonal KO cells were generated by electroporation of multiplexed sgRNA against TRIM34 and single cell sorting (ICE KO score = 96%, Additional file 1: Chromatogram S1). THP-1 TRIM34 KO cells were transduced with doxycycline-inducible HA-tagged primate TRIM34 orthologues or empty vector control. Primate TRIM34 expression was induced in the presence of 125 ng/mL doxycycline, and expression levels were visualized by immunoblotting 72 h post-induction. b-f. Primate TRIM34 expression was induced in THP-1 TRIM34 clonal KO cells. 1 day post-induction, cells were either challenged with chimeric virus particles containing HIV-1 capsids co-expressing zsGreen (b) or SIV capsids co-expressing eGFP including SIVCPZ (c), SIVAGM-SAB (d), or SIVMAC (f); or challenged with VSV-G pseudotyped SIVAGM-TAN-luc (e). Infection was quantified 2 dpi by flow cytometry (b–d, f) or luminometry (e). Relative infectivity in induced cells (white bars, circles) is normalized to average infectivity in uninduced control cells (grey bars, triangles). Fold inhibition is indicated where applicable. Infection of each cell line with each virus was performed 5-6 times across at least 2 different occasions. Combined data are represented as mean +/- s.d., where each point represents a unique infection. One-sided p values were calculated by Welch’s t-test. ns nonsignificant, *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001, ****p ≤ 0.0001