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. 2020 Sep 10;117(39):24415–24426. doi: 10.1073/pnas.2002520117

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Fig. 3. — Concomitant CDK4 inactivation and RAF1 ablation boosts apoptosis. (A) Western blot analysis of CDK4, CDK4^KD, RAF1, phospho-ERK (pERK), ERK1/2 (ERK), phospho-RB (pRB), RB, CDK6, CYCLIN D1, CYCLIN E2, and CDK2 expression in lysates obtained from Kras^+/G12V;Trp53^−/−;hUBC-CreERT2^+/T lung tumor cell lines harboring Cdk4^+/+;Raf1^+/+, Cdk4^FxKD/L;Raf1^+/+, Cdk4^+/+;Raf1^L/L, and Cdk4^FxKD/L;Raf1^L/L alleles 96 h after 4OHT exposure. β-Actin was used as loading control. Migration of the above proteins is indicated by arrowheads. (B) Western blot analysis of CDK4, CDK4^KD, RAF1, cleaved caspase-3 (CC3), caspase-3 (C3) expression in lysates obtained from Kras^+/G12V;Trp53^−/−;hUBC-CreERT2^+/T lung tumor cell lines harboring Cdk4^+/+;Raf1^+/+, Cdk4^FxKD/L;Raf1^+/+, Cdk4^+/+;Raf1^L/L, and Cdk4^FxKD/L;Raf1^L/L alleles maintained in 4OHT containing media. Samples were harvested at the indicated times (hr, hours). β-Actin was used as loading control. Migration of the above proteins is indicated by arrowheads.