Figure 6: Particulate Cr(VI) Causes SAC Bypass not Elimination. This figure shows the effects of triggering the SAC on zinc chromate effects on Cdc20 kinetochore localization and expression levels, and Mad2 protein expression levels. A) One hour before the end of zinc chromate treatment, cells were treated with 0.1ug/ml demecolchicine. Demecolchicine reversed zinc chromate effects on localization. Cells were fixed after the treatment, followed by immunostaining using anti-Cdc20 and centromere antibodies. B) Sixteen hours before the end of zinc chromate treatment, cells were treated with 0.4 ug/ ml nocodazole. Nocodazole had no effect on zinc chromate-induced decreases in Cdc20 protein levels in mitotic cells. C) Representative western blot image of Cdc20 expression in nocodazole co-treated cells. D) Nocodazole had no effect on zinc chromate-induced decreases in Mad2 protein levels in mitotic cells. E) Representative western blot image of Mad2 expression in nocodazole co-treated cells. Protein expression was measured with western blot using anti-Cdc20 and Mad2 antibodies. GAPDH was used as a loading control. Data represent an average of three independent experiments ± the standard error of the mean. *Statistically different from control (p<0.05). †Statistically different from control (p<0.01).