P276-00 on positive regulators of cell cycle along and anti-apoptotic protein: Western blot analysis of cell cycle proteins at designated time intervals after treatment of (A) Jeko-1, (B) Mino and (C) Rec-1 cells with P276-00. Jeko-1 and Mino cells (1-control, 2- IC50 treated and 3- 3X IC50) were incubated with 0.3 and 1 μmol/L P276-00 followed by protein isolation for Western blotting. Rec-1 cells (1-control and 2- treated) were treated at 1.5 μmol/L (3X IC50) of P276-00 and it showed marked decrease in all cell cycle related protein level in time dependent manner. Marked down regulation of anti apoptotic protein, Mcl-1, were also seen from early time point in all three cell lines. Densitometric analysis of cyclin D1 expression was done using ImageJ software. (D) Regulation of cyclin D1 protein levels and Rb phosphorylation by P276-00 was confirmed by immunofluorescence in Mino after 6 h of treatment. Blue: DAPI (nuclear stain); Green: cyclin D1; Red: pRbSer780.