Fig. 2
From: RNA-binding activity of TRIM25 is mediated by its PRY/SPRY domain and is required for ubiquitination
T7-TRIM25ΔRBD supports TRIM25 dimerization. a Co-immunoprecipitation (co-IP) between T7-tagged TRIM25 and EGFP-tagged TRIM25 in cell extracts prepared from HeLa TRIM25 KO cells overexpressing wild-type or ΔRBD TRIM25 mutants. Lanes 1–4 represent loading controls. Lanes 5–8 show co-IPs with anti-T7 antibody. Lanes 9–12 represent control co-IPs with IgG antibody bound to protein-A beads. The bound proteins were analyzed by western blotting with anti-TRIM25 antibody. b Control co-IP shows that co-IP of EGFP-TRIM25 and EGFP-TRIM25 did not occur with anti-T7 antibody. Lanes 1–4 represent loading controls. Lanes 5–8 show co-IPs with anti-T7 antibody. Lanes 9–12 represent control co-IPs with IgG antibody bound to protein-A beads. The bound proteins were analyzed by western blotting with anti-TRIM25 antibody. c Control co-IP in HeLa TRIM25 KO cell extracts reveals that T7-TRIM25ΔRBD and T7-TRIM25ΔPRY/SPRY, but not T7-TRIM25ΔCC or T7-TRIM25-PRY/SPRY, support dimerization. Lanes 1–4 represent loading controls. Lanes 5–8 show co-IPs with anti-T7 antibody. Lanes 9–12 represent control co-IPs with IgG antibody bound to protein-A beads. The bound proteins were analyzed by western blotting with anti-TRIM25 and anti-T7 antibodies