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. Author manuscript; available in PMC: 2023 May 16.
Published in final edited form as: Cell Rep. 2023 Feb 22;42(3):112129. doi: 10.1016/j.celrep.2023.112129

Figure 3. Suppression of E2F phenocopies in deep senescence.

Figure 3.

(A) qPCR of relative E2F1 mRNAs in cells infected with two different E2F1 shRNAs. shGFP was used as control; n=3 biological replicates. Data are represented as mean ± SD; *p < 0.05 by Student’s t test.

(B) E2F1 suppression induces senescence. E2F1-KD A549 cells were stained for β-gal activity. Left: photomicrographs of β-gal-stained control and E2F1-KD cells. Scale bar: 50 μm. Right: percentage of β-gal-positive cells for each condition; n=3 biological replicates. Data are shown as mean ± SD; **p < 0.01 and ***p < 0.001 by two-tailed unpaired Student’s t test.

(C) E2F1 knockdown increases multinucleated cells irrespective of oxygen concentration. Control and E2F1-KD A549 cells were stained with DAPI (red) and tubulin (green). Left: photomicrographs of cells cultured under hypoxia and normoxia. Higher magnification of highlighted multinucleated cells is shown on the right. Scale bars: 50 μm (low magnification) and 20 μm (high magnification). Representative data from two biological replicates are shown.

(D and E) Bar graphs show percentage of binucleated (D) and multinucleated (E) E2F-KD A549 cells grown under hypoxia and normoxia. shGFP is shown as control; n=3 biological replicates. Mean ± SD was calculated from imaging 10 random fields. *p < 0.05 by two-tailed unpaired Student’s t test; n.s, not significant. See also Table S3.