Figure 1. CDK4/6i treatment reduces Rb-protein levels.
(A) Immunoblot showing Rb expression in breast cancer cell lines. Numbers represent relative Rb levels normalized by corresponding GAPDH levels and Rb levels in MCF-7 cells.
(B and D) Density scatterplots of Hoechst and EdU staining in Rb-mutant MDA-MB-468 (B) and MCF-7 (D) cells treated with palbociclib (1 μM) (n = 2,000 cells/condition).
(C and E) Percentage of S-phase cells after palbociclib (1 μM) treatment in breast cancer cells. Data are shown as means ± SD (n = 3 biological replicates).
(F) Immunoblot showing Rb expression in MCF-7 and MDA-MB-231 cells treated with palbociclib (1 μM).
(G) Percentage of S-phase cells after palbociclib (1 μM) treatment in Rb-mutant MDA-MB-468 cells with or without YFP-Rb overexpression. Data are shown as means ± SD (n = 3 biological replicates).
(H) Density scatterplots of Hoechst and EdU staining in MCF-7 cells overexpressing YFP-Rb and treated with palbociclib (1 μM) (n = 2,000 cells/condition).
(I) Percentage of S-phase cells after palbociclib (1 μM) treatment in MCF-7 and MDA-MB-231 cells with or without YFP-Rb overexpression. Data are shown as means ± SD (n = 3 biological replicates).
(J) Immunoblot showing expression of endogenous and exogenous Rb in MCF-7 and MDA-MB-231 cells treated with palbociclib (1 μM).
(K) Relative levels of endogenous and exogenous Rb protein in MCF-7 cells treated with palbociclib (1 μM). Solid lines indicate best fitted lines. The half-life of Rb protein is shown as means ± SD (n = 3 biological replicates).
(L) Relative mRNA levels of Rb and E2F target genes in MCF-7 cells treated with palbociclib (1 μM). Data are shown as means ± SD (n = 3 biological replicates).
See also Figures S1 and S2.