Figure 3: Basal p21 expression correlates with the presence of DNA damage foci. | Nature Communications

Figure 3: Basal p21 expression correlates with the presence of DNA damage foci.

From: DNA damage during S-phase mediates the proliferation-quiescence decision in the subsequent G1 via p21 expression

Figure 3

(a) Images of hTert-RPE1 mRuby-PCNA p21-GFP cells fixed and stained for γH2AX. Magnified regions of each image are also shown. p21-GFP is green, γH2AX is red in merged images. Scale bars are 10 μm. (b) Quantification of p21-GFP nuclear fluorescence in interphase (excluding S-phase) cells with or without γH2AX nuclear foci. With foci, n=2086 cells; without foci, n=12476. (c) Quantification of number of γH2AX foci per nucleus after separation of cells based on a high or low p21-GFP nuclear fluorescence (high p21>30 a.u.). High p21-GFP, n=3896 cells; low p21-GFP, n=11049. (d) Fates of sister cells pairs (D1 and D2) with regards to 53BP1 focus appearance. 142 daughter pairs were analysed. (e) Correlation between the time of 53BP1 focus appearance in sister cells. 45 sister pairs are shown. Note: 14 sister pairs are clustered at time 0,0 min and are not visible as separate values. Pearson’s Correlation R=0.39* (P<0.05). (f) Histogram of G1 length separated by the presence or absence of a 53BP1-GFP focus. The difference between the two populations is significant (unpaired student’s t-test P=0.0003). 118 cells were scored manually.

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