SUPPLEMENTAL TABLE 1. Cell cycle profiles of HeLa cells treated TAB. S1. Leung-Pineda et al. SUPPLEMENTAL TABLE 1. Cell cycle profiles of HeLa cells treated with HU, Gö6976 or OA Table S1. Short-term treatment of HeLa cells with Gö6976 or OA does not alter cell cycle profiles. HeLa cells were left untreated (Control) or were incubated with 1 M OA for 1 h , 1 M Gö6976 for 1 h or 20 mM HU for 2 h. Alternatively, HeLa cells were incubated with 20 mM HU for 2 h, the culture media was removed and cells were incubated in complete media containing DMSO or 1 M OA or 1 M Gö6976 for 1 h. Cells were stained with propidium iodide and analyzed for DNA content by flow cytometry. The percentage of cells in each stage of the cell cycle is indicated.
A B FIG. S1. Leung-Pineda et al. OA Control DMSO Concentration (M) : HU 0.03 0.3 1 5 0.05 0.5 1 2 pS345-Chk1 pS317-Chk1 Chk1 1 2 3 4 5 6 7 8 9 10 11 B DMSO Gö6976 (1 M) OA (1 M) Control Time (min) : HU 15 30 45 60 90 15 30 45 60 90 15 30 45 60 90 pS345-Chk1 Chk1 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 Figure S1. Dose dependent inhibition of Chk1 dephosphorylation by Gö6976 and OA. (A) HeLa cells were untreated (lane 1) or were incubated with 20 mM HU for 2 h to generate phosphorylated Chk1 (lanes 2-11). The culture media was removed and cells were incubated in complete media containing DMSO (lane 3) or the indicated concentrations of either Gö6976 (lanes 3-7) or OA (lanes 8-11). Chk1 phosphorylation was monitored 1 hour after HU removal by Western blotting. (B) HeLa cells were treated as described in panel A except that the concentration of Gö6976 and OA were held constant and the time of incubation was varied.