To understand in spanish5/7/2023 ![]() ![]() The experiment was performed twice with similar results. The immunoblot on the left shows remaining protein levels in total cell extracts in the different conditions. d Flow cytometry contour plots for chromatin-bound STAG1 and STAG2 in control (gray contour plots), SMC1 KD and double NIPBL/SMC1A KD (colored contour plots) HeLa cells. A non-parametric Mann–Whitney two-sided test with confidence intervals of 99% was performed. At least 372 cells were analyzed per condition in a single experiment. units, arbitrary units) of control or NIPBL KD HeLa cells stained with antibodies against STAG1, STAG2 and SMC1A. This is one representative experiment of at least 3 performed. NIPBL partner MAU2 also decreases after NIPBL KD. Increasing amounts of total extract from control cells were loaded to better quantitate the extent of depletion. b Immunoblot analysis of chromatin fractions (Chr) and total cell extracts from control and NIPBL KD cells. For each map, the cell cycle profile according to DNA content appears on top while the distribution of antibody intensities is plotted on the right. Contour plots of the indicated proteins in control (gray plots) and NIPBL KD cells (colored plots) were overlapped for comparison. a Asynchronously growing HeLa cells mock transfected (control) or transfected with siRNA against NIPBL (NIPBL KD) were analyzed by flow cytometry 72 h later. NIPBL KD affects cohesin-STAG1 and cohesin-STAG2 in opposite ways. ![]()
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