Fig. 7.

Cells partially depleted of SMC2 show chromosome mis-segregation. (A) Experimental timeline for auxin treatment. Auxin was omitted from the medium used for the wash and release except in the case of cells treated with 125 or 250 µM auxin. (B,C) Flow cytometry analysis of SMC2-AID/CDK1^as cells. Cells were analyzed at the end of 13 h 1NMPP1 treatment (0 min, G2) and 30 min after washout of 1NMPP1 (30 min, mitosis) with various concentrations of auxin treatment followed by washout. The relative median of the GFP signal is shown (C). (D) The amount of SMC2 remaining in asynchronous culture upon 4 h auxin treatment was measured by immunoblot analysis and normalized to the level of α-tubulin. Data are plotted as mean±s.d. (n=3). (E) Representative images of SMC2-AID/CDK1^as cells at prometaphase after a 4 h treatment with various concentrations of auxin. Scale bar: 5 µm. (F) Effect of various auxin concentrations on mitotic progression of SMC2-AID/CDK1^as cells. After release from 4 h 1NMPP1 treatment, cells were fixed every 30 min. >200 cells were counted for each time point of individual treatments. Data are plotted as mean±s.d. (n=3).