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

Figure 2. Roscovitine enhances the expression of ATRA-induced enrichment of nuclear Src-family kinase members.

Figure 2

Nuclear lysates collected after 72 h of treatment were resolved on 12% polyacrylamide gels. 25 μg protein was loaded per well. (A–C) Roscovitine enhances ATRA-induced nuclear Lyn, Fgr and Y416-c-Src expression. p < 0.05 comparing ATRA-treated samples to ATRA/roscovitine-treated samples. (D) TATA binding protein (TBP) was the loading control. (E) Roscovitine augments ATRA-induced reduction of nuclear Lyn interaction with pS608 phosphorylated RB tumor suppressor protein. Co-immunoprecipitation was done using Lyn as bait. (F) Nuclear RB binds Lyn in ATRA and ATRA plus roscovitine treated cells. Co-immunoprecipitation was done in treated cells using RB as bait. Vav also binds RB in these cells. An equal amount of pre-cleared nuclear lysate was collected 72 h post treatment and incubated overnight with 1:100 concentration of the precipitating antibody with magnetic beads and resolved on 12 % polyacrylamide gels. All blots shown are representative of three replicates.