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The effect of siRNA depletion of Tim was first examined by cell cycle profile using FACS analysis of propidium iodide stained cells, 24 hrs post-transfection (Fig. 3B).
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Therefore, we concluded that irradiation did not potentiate ONYX-015 by changing the cell cycle profile.
Cell growth inhibition was confirmed by analyzing the cell cycle profile for each cell line after 72 h of treatment.
We tested this hypothesis by analyzing the cell cycle profile following siRNA-mediated depletion of Smek2, PHF6 and FUBP1 (Fig 9A).
Using a matrix population model (see " Methods"), we computed that the slower reproductive schedule imparted by the optimal cell cycle profile derived from optimization 5 leads to a sixfold increase in population doubling time compared to a flat cell cycle length profile fixed at 2.8 h (76 h vs. 12 h, respectively).
Therefore, the higher apoptosis induced by Aza-dC treatment in control than in DNMT3B knockdown cells, which was observed only in NTERA2 but not other cell lines, could be explained by the formation of covalent adduct between Aza-dC-DNMT3B at genomic level rather than by difference in cell cycle profile especially in S phase.
Data were acquired by Flow Cytometry and cell cycle profile was processed by FlowJo software (Tree Star Inc, Ashland, OR, USA).
The effect of SAHA on blocking cell proliferation through blocking DNA synthesis was examined by BrdU labeling, as determined by immunofluorescence staining (Fig. 2A), and the cell cycle profile by flow cytometric analysis (Fig. 2B).
To investigate the mechanism by which TS depletion inhibits lung cancer cell growth, we examined the cell cycle profile by flow cytometry.
Next, we tested cell cycle profile by DNA staining with propidium iodide (PI), followed by flow cytometry analysis.
We also analyzed the cell cycle profile by propidium iodide (PI) staining on day 3 and day 4.
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