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. 2010 Sep 2;5(9):e12532. doi: 10.1371/journal.pone.0012532

Figure 5. Regulation of cell cycle-controlling genes following Bcl11b accumulation.

Figure 5

(A) The protein lysates acquired from mock- and BCL11B-transduced Jurkat and huT-78 cells were analyzed by Western blotting using indicated antibodies 48h post-transduction. (B) The effects of BCL11B overexpression on ubiquitin ligase SKP2 was evaluated by semi-quantitative RT-PCR and western blot 48h after transduction. (C) The retinoblastoma protein phosphorylation status was assayed using antibodies preferentially binding to hypophosphorylated variant of pRb followed by flow cytometry. Additionally, the antibodies recognizing specifically the phosphorylated pRb and total retinoblastoma-like protein 2 were used to detect the proteins by western blot. (D) The amount of Myc-N protein in cells with normal and elevated Bcl11b levels were assessed by immunodetection in protein lysates (upper part, western blot). In addition, a Myc-N-specific antibody was used for intracellular staining followed by flow cytometry (lower panel, density plot shows reporter gene – X axis, and Myc-N – Y axis). All western blot and FACS data shown on this figure represent results obtained from at least three experiments. Section B (upper part, semi-quantitative RealTime RT-PCR) shows average values from three experiments ± SD, *p<0.05.