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. Author manuscript; available in PMC: 2011 Feb 15.
Published in final edited form as: Clin Cancer Res. 2010 Feb 9;16(4):1179. doi: 10.1158/1078-0432.CCR-09-1787

Figure 5. Roscovitine blocks the AD-induced increase in active (phosphorylated) CDK2 and LMW cyclin E overexpression cannot bypass this effect.

Figure 5

Figure 5

(A) Schematic representation of the experimental design. (B) MCF-7/Ac1 cells were cultured in IMEM with 10% charcoal-stripped serum medium (CSSM) without phenol red and with 600 µg/ml of G418 for 4 days before plating at a density of 100,000 cells for a 100 mm dish. Cells were then infected with the indicated adenoviruses (at 4000 m.o.i.) 24 hours before drug treatment. Cells were then treated with 25 nM AD and 1 µM letrozole and collected 3 days later for cell number (3d) followed by 2 days in medium (CSSM) alone (3d + 2d) or medium (CSSM) plus DMSO (3d + 2d DMSO) or medium (CSSM) plus 20 µM of roscovitine (3d + 2d Rosco). (C) The same cells used for counting were collected and lysates were subjected to western blot analysis (25 µg of protein) with either the CDK2 (D-12) or phospho-T160-CDK2 antibody. (D) upper, Ratio of densitometric values of phosphorylated CDK2/total CDK2. Lower, Densitomertic values of the phospho-T160-CDK2 bands in AD + Let treated cells.