Figure 5
Figure 5. Effects of TIP110 expression on differentiation, cell-cycle status, and apoptosis of CD34+ cells. Freshly isolated CD34+ cells were immediately transfected with expression plasmids for GFP, TIP110.GFP, shRNA control (shCtrl), or shRNA for TIP110 (shTIP110). These cells were cultured in IMDM with 1% FBS without the addition of any growth factors for 2 days, then harvested for TIP110 mRNA expression by semiquantitative RT-PCR (A), percentage of CD34-expressing cells by FACS (B), and cell-cycle analysis and apoptosis by flow cytometry for TIP110 (C-D). Shown are GFP-overexpressing (C) and shTIP110-underexpressing (D) CD34+ cells. Data are shown as means ± SEM for 3 experiments, each done in triplicate. K562 cells were transiently transfected with expression plasmids for GFP, TIP110.GFP, shRNA control (shCtrl), or shRNA for TIP110 (shTIP110). Cells were cultured for 48 hours and harvested for evaluation of TIP110 expression by Western blotting (E) or assessed by the MTT assay for cell proliferation (F). Data in panels E and F are from 1 of at least 3 reproducible experiments.

Effects of TIP110 expression on differentiation, cell-cycle status, and apoptosis of CD34+ cells. Freshly isolated CD34+ cells were immediately transfected with expression plasmids for GFP, TIP110.GFP, shRNA control (shCtrl), or shRNA for TIP110 (shTIP110). These cells were cultured in IMDM with 1% FBS without the addition of any growth factors for 2 days, then harvested for TIP110 mRNA expression by semiquantitative RT-PCR (A), percentage of CD34-expressing cells by FACS (B), and cell-cycle analysis and apoptosis by flow cytometry for TIP110 (C-D). Shown are GFP-overexpressing (C) and shTIP110-underexpressing (D) CD34+ cells. Data are shown as means ± SEM for 3 experiments, each done in triplicate. K562 cells were transiently transfected with expression plasmids for GFP, TIP110.GFP, shRNA control (shCtrl), or shRNA for TIP110 (shTIP110). Cells were cultured for 48 hours and harvested for evaluation of TIP110 expression by Western blotting (E) or assessed by the MTT assay for cell proliferation (F). Data in panels E and F are from 1 of at least 3 reproducible experiments.

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