Figure 2

Cell cycle-dependent hMSH5 expression and cisplatin-triggered hMSH5 protein induction. (A) FACS analysis of cell cycle synchronization. Data were analyzed by the use of FlowJo software (Dean-Jett model). Values are means ± standard deviation from 3 independent measurements. The expression of hMSH5 in 293T cells synchronized to G1, S, G2/M phases, and asynchronized controls were examined by Western blot analysis. α-Tubulin immunoblot was used as a loading control. (B) Cisplatin-triggered hMSH5 induction in S phase cells. 293T cells were synchronized at S phase and treated with 10 μM cisplatin for 8 hrs and followed by Western blot analysis of hMSH5 protein levels. α-Tubulin immunoblot was used as a loading control. (C) Treatment of S phase cells with cisplatin enhanced hMSH5 nuclear accumulation. Cells synchronized at S phase were treated with 10 μM cisplatin for 8 hrs, and hMSH5 protein levels in the nuclear and cytoplasmic fractions were evaluated by Western blotting. RNAPII and α-tubulin were used as nuclear and cytoplasmic loading controls, respectively.