FIGURE 6.

Effect on cell cycle by SUMO-regulated phosphorylation. A, proteins involved in cell cycle control were enriched in up- and down-regulated groups. B, HEK293T cells were treated with DMSO and 100 μm ginkgolic acid for 6 h before harvest, then lysed, and blotted (IB) with anti-Cdc2 and anti-pTyr Cdc2 antibody. Actin was used as a loading control. C, flow cytometry analysis of the cell cycle upon ginkgolic acid treatment. Representative original flow cytometry data are shown on the right. HEK 293T cells were treated with DMSO or 100 μm ginkgolic acid for 1 h, 4 h, 6 h, and 8 h. Cells were harvested, stained with propidium iodide, and analyzed by flow cytometry. n = 3; *, p < 0.05; **, p < 0.01. D, flow cytometry analysis of the cell cycle after knockdown of SUMO expression. Representative original flow cytometry data are shown on the right. HEK 293T cells were transfected with shRNA plasmids against LacZ control, SUMO1, or SUMO2/3 for 48 h. Cells were harvested, stained with propidium iodide, and analyzed by flow cytometry for cell cycle changes. n = 6; **, p < 0.01. All the original flow cytometry analysis of cell cycle were included in Supplemental Fig. S3.