Fig. 3

Different chromatin association dynamics of the two cohesin variants. a Chromatin fractions isolated from MEFs of the indicated genotypes were challenged or not with 0.5 M NaCl and the amount of protein remaining on chromatin was assayed by immunoblot. b Quantification of the fraction of RAD21 left on chromatin after salt extraction (4 replicates). c iFRAP analyses of G0 MEFs expressing STAG1-GFP, STAG2-GFP or RAD21-GFP. Curve represents the difference in mean fluorescence intensity between the bleached and unbleached areas as a function of recovery time (min). Imaged cells were n = 20 for RAD21-GFP, n = 24 for STAG1-GFP and n = 16 for STAG2-GFP expressing cells from 4 independent experiments. Images from videos at the indicated timepoints are shown below. Bar, 5 µm. d Immunoprecipitation reactions from iMEF extract with the indicated antibodies (or IgG as control) were analyzed by western blot. e. Amount of WAPL pulled down relative to RAD21 was quantified in three independent experiments including the one shown in d