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. 2024 Feb 22;4(3):319–335. doi: 10.1038/s43587-024-00575-6

Fig. 8. Local PS externalization is associated with PMD-dependent and replicative senescence.

Fig. 8

a, SA-β-gal (green) and Annexin V–Alexa Fluor 488 (red) signals were observed in HCA2 senescent cells induced by three different senescence induction methods (6 d after SDS wash away (SDS) and RS (cell division) and 6 d after IR). Scale bar, 50 μm. White arrows, an Annexin V–positive projection. This experiment was independently repeated three times with similar results. b, Rep-Sen WI-38 cells were incubated with Annexin V–Alexa Fluor 647 conjugate, fixed and stained with CHMP4A antibody. The white rectangle region in the left panels is enlarged in the right panels. Green, CHMP4A; red, Annexin V–Alexa Fluor 647 conjugate. BF, bright-field. White arrows, Annexin V and CHMP4A co-localization. Scale bar, left: 10 μm, right: 2 μm. This experiment was independently repeated three times with similar results. c, HCA2 cells were treated with 0.01% SDS for 24 h, washed and released into fresh medium. The cells were fixed after 6 d. SDS-treated (SDS, senescent) and untreated (Control, young) cells were analyzed by FIB-SEM. Scale bar, 1 μm. This experiment was independently repeated three times with similar results.