Figure 3.
PRMT5 accelerates progression through G1 and upregulates regulators of G1 phase. (a) The EGFP and PRMT5 clonal cell lines were analyzed by flow cytometry, and the percentage of cells in each specific phase was calculated and plotted. (b) The protein level of various cell cycle regulators in the EGFP and PRMT5 clonal cell lines was analyzed by western blotting. Antibodies against cyclins A, B1, D1–D3, E1, and E2; CDK2, 4, and 6; p19; Rb protein; phospho‐Rb protein; and β‐actin were used for western blotting. (c) The kinase activities of various CDKs were assayed by performing in vitro kinase reaction where CDK2, CDK4 or CDK6 was isolated by immunoprecipitation employing specific antibodies from cells harboring nothing, EGFP empty vector, EGFP‐PRMT5, PRMT5 shRNA, or scrambled shRNA. Recombinant Histone H1 or Rb was used as substrate and incubated with CDKs in kinase reaction buffer in the presence of [γ‐P 32]‐ATP. *Statistical significance by Student's t‐test with P < 0.05.