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. 2022 Jan 12;2:797562. doi: 10.3389/fragi.2021.797562

FIGURE 3.

FIGURE 3

A G4-stabilizing compound exacerbated senescence-associated phenotypes in cultured CEC. (A) CEC isolated from aged (18–20 m/o) female mice and cultured were treated with pyridostatin (0.5 µM), or a vehicle (water), for 4 days. CEC were fixed and stained with antibodies against γH2AX and the nuclear Hoechst dye (DAPI channel). Scale bar, 50 µm (B) Quantification of the fluorescence intensity of γH2AX from (A). t-test, ***p-value = 0.0003. Data were pooled from three independent experiments analyzing 50 cells per condition and experiment. (C) Quantification of the nuclei size. t-test, ***p-value = 0.0001. Data were pooled from three independent experiments analyzing 50 cells per condition and experiment. (D) CEC isolated from aged (18–20 m/o) female mice and cultured were treated with pyridostatin (0.5 µM), or a vehicle (water), for 4 days. CEC were fixed and stained with the SA-β-galactosidase staining kit. Scale bar, 50 µm. (E) Quantification of the percentage of SA-β-galactosidase-positive cells. t-test, **p-value = 0.0015. Data were pooled from three independent experiments analyzing 50 cells (A–C) or 20 microscopic fields (D,E) per condition and experiment.