Fig. 5.

RSF1-deficient cells suffering from sister chromatid cohesion defects are rescued by centromere targeting HDAC1. a RSF1 KO HeLa cells were transfected with centromere targeting GFP-CENP B fused-HDAC1 WT or H141A catalytic dead mutant constructs. Floating mitotic cells were obtained after nocodazole treatment for 4 h and subjected to chromosome spread immunostaining with anti-H2A-pT120 (red) antibodies. Scale bar, 5 μm. b The cells prepared as Fig. 5a were subjected to chromosome spread immunostaining with anti-Sgo1 (red), and DAPI (blue). Scale bar, 5 μm. c Immunoblotting of chromatin fractions in RSF1 KO cells after reintroduction of CENP B fused GFP-HDAC1. d Giemsa staining of the metaphase chromosome spreads prepared after transfection of GFP-CENP B fused HDAC1 WT or H141A in RSF1 KO cells. Left panel shows quantification of the percentage of premature sister chromatid separation