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Representative images of DNA fibers of TKO-Bcl2 MEFs with and without 10% FCS (lower panel).
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Ongoing forks were used to determine fork speeds (kb/min) 1 st label and 2 nd label origins are origins of replication initiated during the labelling period with CldU and IdU, respectively (upper panel). ( A) Schematic representation of replication tracks generated after pulse labeling with CldU (red) and IdU (green). Experiments in A, B and E were performed in triplicate. ( E) IncuCyte growth curves of TKO-Bcl2 (black), TKO-Bcl2-p53KO (red) and TKO-Bcl2-p21KO (blue) MEFs in the absence of 10% FCS. Percentage of BrdU-labeled cells is indicated. ( D) BrdU flow cytometry analysis of the cell cycle distribution of TKO-Bcl2 and TKO-Bcl2-p53KO MEFs in the absence of 10% FCS for the indicated days. ( C) Cell cycle distribution based on propidium iodide content of TKO-Bcl2 MEFs (upper panel) and TKO-Bcl2-p53KO MEFs (lower panel) in the absence of 10% FCS for the indicated days. Apoptosis was measured by fluorescent signal upon caspase three cleavage and normalized to cell confluency. ( B) Apoptosis levels of TKO-Bcl2 (black), TKO-p53RNAi (green), TKO-p53KO (blue) and TKO-Bcl2-p53KO (red) MEFs in the absence of 10% FCS. ( A) IncuCyte growth curves of TKO-Bcl2 (black), TKO-p53RNAi (green), TKO-p53KO (blue) and TKO-Bcl2-p53KO (red) MEFs in the absence of 10% FCS.