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Fig. 2 | SpringerPlus

Fig. 2

From: ZD7288, a blocker of the HCN channel family, increases doubling time of mouse embryonic stem cells and modulates differentiation outcomes in a context-dependent manner

Fig. 2

S-phase lengthening is not mediated by increased gammaH2AX or altered S-phase progression, but is accompanied by reduced nucleoside incorporation. Control cells and cells treated with ZD7288 for 5 days were stained for gammaH2AX (a), with no difference in staining intensity (Student’s t-test; not significant; n = 3) (b). Cells grown in bulk with and without the addition of ZD7288 were pulsed with 10 μM EdU prior to fixation and stained for flow cytometric analysis. Treated cells incorporated less EdU (p < 0.05, n = 3) (c). Cells grown adherently for 4 days in control or ZD7288 conditions were also pulsed and stained for EdU (d), confirming lower levels of incorporation. Cells with EdU incorporation pattern typical of early S-phase (d, star and inset) and mid-late S-phase (d, arrowhead and inset) in both conditions were counted (e) with similar distribution of cells between early and late S under both conditions (Student’s t-test; not significant; n = 3)

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