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Fig. 4 | Genome Medicine

Fig. 4

From: Functional fingerprinting of human mesenchymal stem cells using high-throughput RNAi screening

Fig. 4

PIK3C2A and WEE1 silencing alters the cell cycle profile of primary MSCs. Cytometric analysis of MSCs 96 h after siRNA treatment confirms viability phenotypes of selected candidates. a Representative fluorescence flow cytometry analysis of different MSC donors. MSCs were stained with 200 μg/ml of propidium iodide (PI), 0.1 %, phycoerythrin (PE), 0.1 %, sodium azide, 0.1 % Triton-X100 and 10 μg/ml RNAses for 2–4 h at 4 °C. Single cells were analyzed for fragmented DNA and dead cells were gated to quantify viable cells (PE-A). The percentage of viable cells was calculated from three biological replicates and average viability was plotted. b Representative histograms of MSCs analyzed with FlowJo 887 for sub G1, S and G2 peaks (PI-A). PIK3C2A and WEE1 show a shift in DNA content (blue) compared with the negative control (red). Quantification of cell cycle percentages shows that PIK3C2A and WEE1 silencing led to a significant (p ≤ 0.01) increase in S-phase and G2 peaks, respectively. Average of three biological replicates is shown and significant cell cycle changes were calculated using unpaired two tailed student’s T-test

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