NBR1 F740A and TAX1BP1 R441E are sufficient to disrupt in vivo correlation. (A) Representative confocal micrographs of Hela PentaKO cells co-transduced with tf-NBR1 or tf-NBR1F740A and BFP-TAX1BP1 or BFP-TAX1BP1R441E. Scale bar: 10 µm. (B) Quantification of A. Data represent median of three independently generated cell lines. Pearson’s correlation coefficient between RFP and BFP (reflective of NBR1 and TAX1BP1 co-localization) was calculated using Cell Profiler. Scatter plots represent median ± SD. Each data point represents a single replicate. Statistical analysis was performed using ordinary one-way ANOVA with Tukey’s multiple comparisons correction. n ≥ 17 cells per replicate; ****, P < 0.0001, ns, not significant. (C) HEK293T cells were co-transfected with full-length (FL) myc-TAX1BP1 and indicated BFP-V5-TAX1BP1 variants. Extracts derived from transfected cells were immunoprecipitated (IP) with protein G dynabeads conjugated with anti-V5 antibody. Input and eluates were resolved by SDS-PAGE followed by immunoblotting (IB) with the indicated antibodies. Source data are available for this figure: SourceData FS6.