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. 2020 Mar 24;11(12):1017–1036. doi: 10.18632/oncotarget.27508

Figure 6. Roscovitine effects on ATRA-induced changes in nuclear expression of G1/G0 regulatory molecules: p27/cyclin E1/Cdk2/RB pathway.

Figure 6

(A) Western blot of p27Kip1 in cells cultured for 72 h showed that ATRA enhanced nuclear p27Kip1 level, and cells co-treated with ATRA and roscovitine modestly further upregulated the p27Kip1 expression. (B) Roscovitine further decreased ATRA-induced reduction of nuclear cyclin E1 expression in these cells. (C–E) Roscovitine diminishes ATRA-induced downregulation of nuclear CDK2 and specifically its pY15CDK2 and pT160CDK2 phosphorylated forms. (F, G) ATRA plus roscovitine downregulates total RB and pS608 phosphorylated RB. Roscovitine enhanced ATRA-induced reduction of RB phosphorylated at pS608 site. Surprisingly, at the same dose, roscovitine alone enhances nuclear cyclin E1 and CDK2 expression. p < 0.05 comparing ATRA-treated samples to ATRA/Roscovitine-treated samples. (H) TATA binding protein (TBP) was the loading control; a minor artifact caused during image capture can be seen. All blots shown are representative of three replicates.