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. 2009 Jul 27;284(38):25664–25677. doi: 10.1074/jbc.M109.014241

FIGURE 3.

FIGURE 3.

c-Cbl G306E failed to promote RA-induced cell cycle arrest. A, representative DNA histograms of untreated and (72 h) RA-treated vector control, c-Cbl+, G306E, and C381A stable transfectants. B, percentage of vector control, c-Cbl+, G306E, and C381A transfectant cells with G1/G0 DNA after a 72-h RA treatment. Cells stained with hypotonic propidium iodine (PI) staining solution were analyzed by flow cytometry. Bars represent means ± S.E. of 8 repeats. The different letters indicate significant differences at the p ≤ 0.05 levels between untreated (C) and RA treated (RA) among the same cell lines. Asterisk indicates a significant (p ≤ 0.05) difference compared with vector control cells. C, Western blots of CDK4 for vector control, c-Cbl+, G306E, and C381A stable transfectants. The WT c-Cbl and the C381A transfectants had less CDK4 expression than vector control or G306E transfectants after RA treatment (72 h). GAPDH was used to check protein loading. The data from the CDK4 Western blots were consistent with G1/0 arrest assays, indicating that the G306E mutant failed to promote cell cycle arrest.