Neither A216 nor L217F mutants of PML recruit p97 to nuclear bodies upon arsenic treatment. (A) Representative fluorescence microscopy images for 1 μM arsenic-treated YFP-PML-V cells at 0 and 6 h, showing YFP fluorescence and p97 immunofluorescence. (B) Quantitative summary of p97 colocalization with PML bodies in the indicated cell lines during 0-, 2-, 4-, and 6-h arsenic exposure. Markers show average values, and error bars are SEM. (C) Statistical summary of the comparison between 0- and 6-h arsenic exposure for YFP-PML-p97 colocalization. Columns represent average % colocalization per cell, and error bars are SEM. Cell count (n) is shown in brackets below each column. Statistical significance was assessed by comparing the arsenic-treated conditions among the three PML types using Dunn’s multiple comparisons tests after Kruskal–Wallis ANOVA. (D) Anti-PML and anti-p97 immunoblots of crude cell extracts taken from the indicated cell lines either treated or not with 1 µM arsenic for 6 h; WT U2OS (U2OS) or PML^−/− U2OS expressing the YFP-PML-V variants (WT, A216T, or L217F). *Note multiple modified and unmodified endogenous PML isoforms are difficult to distinguish. Source data are available for this figure: SourceData F5.