Extended Data Fig. 2: Further characterisation of TRIM21 RING domain mutants.
From: Target-induced clustering activates Trim-Away of pathogens and proteins
a,b, RING dimerisation mutants M10E and M72E are expressed at WT levels (a) and do not affect AdV5 infection efficiency (b). c, His-Ubiquitin pulldown from cells expressing the indicated TRIM21 mutants infected with AdV5 ± 9C12 or 9C12(H433A). *T21 shows non-specific binding of unmodified TRIM21 to the NiNTA beads. e,f, Second site mutants R67A and N71D are expressed at WT levels (e) and do not affect AdV5 infection efficiency (f). g, Neutralisation of AdV5 infection by increasing 9C12 concentrations in cells expressing the indicated TRIM21 mutants. h, AdV5-9C12-induced NFkB activation in cells expressing the indicated TRIM21 mutants. i, Trim-away of target protein IKKα following electroporation (+) of anti-IKKα IgG (anti-IKKα). Immunoblots show IKKα degradation is deficient in cells expressing the N71D mutant TRIM21. Graphs in b,f,g,h show mean and s.e.m for n = 3 independent experiments. Statistical significance is based on one-way ANOVA (b,f,h) and two-way ANOVA (g) and represented with symbols: ns(P>0.05), *(P≤0.05), **(P≤0.01), ***(P≤0.001), ****(P≤0.0001). Uncropped blots/membranes are in Supplementary Data 2. Source data for graphs and statistics are in Supplementary Data 1.
