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. 2023 Sep 13;51(18):9594–9609. doi: 10.1093/nar/gkad756

Figure 6.

Figure 6.

A conserved FDF motif in PAGR1 promotes cohesin occupancy on chromatin. (A) Sequence alignment of PAGR1 at the FDF motif. Box indicates the consensus FDFDD sequence, blue amino acids indicate phenylalanines mutated to alanines in the ADA mutant. (B, C) PAGR1-KO cells were stably transduced with empty vector (EV), FLAG-tagged WT or mutant (ADA) PAGR1. Whole cell extract (B) and chromatin-bound protein fractions (C) were assessed by western blot. (D) Flag-PAGR1 was immunoprecipitated from indicated cell lines followed by western blot analysis. (E) SMC3 band intensities were quantified from three independent western blots relative to WT-EV levels, corrected for Histone H3. P-values were calculated by one sample T tests against a null hypothesis of 1. (F) RAD21 intensity was quantified in indicated cell lines in three independent experiments with at least 142 cells per sample per experiment. Box represents 25%-75% and median, whiskers represent 1–99 percentile from combined data points. P-values were calculated by a one-way ANOVA comparing the median values from separate biological replicates, which are indicated by white diamonds. Scale bar represents 10 μm.