Fig. 5

PDS5 proteins and STAG proteins have differential effects on SMC3ac levels and chromatin bound cohesin. A Western blot analysis of nuclear lysates from seven siRNA conditions in mESCs. Abbreviations included for PDS5A (P5A), PDS5B (P5B), STAG1 (SA1), and STAG2 (SA2). B Percentage of cells in each cell cycle phase, n = 3. A 2-tailed unpaired t test was used to determine significance. No significance was found between any mutant and WT condition for any cell cycle phase. C Western blot analysis following fractionation of cells from seven siRNA conditions. Both chromatin-bound (B) and nuclear soluble (U) are shown for each condition. D Western blot analysis following co-immunoprecipitation of IgG (negative control), SMC3, and NIPBL in WT siGLO and Pds5a^−/− siPds5b nuclear lysates, under high stringency conditions. Black triangle indicates the NIPBL-specific band (top), as determined via NIPBL knockdown (see Additional file 2: Figure S6D). E ChIP–qPCR for NIPBL in WT siGLO and Pds5a^−/− siPds5b mESCs. Fold enrichment relative to 5% input material and the negative control region is depicted. Data represented as mean ± standard deviation across two biological replicates, each with three technical replicates. A 2-tailed unpaired t test was used to determine significance between WT siGLO and Pds5a^−/− siPds5b mESCs for every primer set. No significance was found for any of the primer sets. F ChIP–qPCR for WAPL in WT siGLO and Pds5a^−/− siPds5b mESCs. Fold enrichment relative to 5% input material and the negative control region is depicted. Data represented as mean ± standard deviation across two biological replicates, each with three technical replicates. A 2-tailed unpaired t test was used to determine significance between WT siGLO and Pds5a^−/− siPds5b mESCs for every primer set. Asterisks indicate significant differences between groups (****p < 0.0001), otherwise, no significance was found