Figure 8.

Cohesin-interacting splicing factors are required for mitotic progression. A, validation of siRNAs targeting representative cohesin-interacting splicing factors in HeLa cells. HeLa cells were transfected with negative control siRNA (s4390843) or gene-specific siRNAs as indicated by the Ambion catalogue number, cultured for 72 h, and harvested in RIPA buffer, and Western blotting was performed with the antibodies indicated. B, HeLa-H2B-GFP cells were transfected with siRNAs for SNRNP200 (ss22846), EFTUD2 (s17862), PRPF31 (s25123), SF3B3 (s23849), and sororin (s535461) and studied by live-cell imaging as described under “Experimental procedures.” Representative aberrant mitoses in siRNA-transfected cells are shown. Bar = 10 μm. For time-lapse movies with images every 5 min from 36 to 72 h post-transfection, see Movies S1 and S2. C, quantification of aberrant mitoses in siRNA-transfected cells. Cells in four fields were counted over the 36-h period of observation for each siRNA. The data shown are from three independent biological replicates. Error bars indicate standard deviation.