Figure 7. Two-species system: knockdown of human SMC3 by siRNA and rescue by stably expressed EGFP-msSMC3.

(A) Cells stably transfected with EGFP-msSMC3 were analyzed by IF microscopy. (B) Total cell (RIPA), cytoplasmic and nuclear extracts from cells stably expressing EGFP-msSMC3 collected 72 h after treatment with siSMC1 or siSMC3 were analyzed by IB using anti-EGFP, -SMC3 and -SMC1 antibodies. RIPA buffer was used for the final extraction step after the nuclear extraction with 250 mM ammonium sulfate. Topo II and Karyopherin ß1 were used as a loading control. Relative protein levels are shown at the bottom (representative of 3 experiments).