Extended Data Fig. 9: Human WI-38 fibroblast senescence validation. | Nature Aging

Extended Data Fig. 9: Human WI-38 fibroblast senescence validation.

From: Single-cell and spatial transcriptomics map senescent vascular cells in arterial remodeling during atherosclerosis in mice

Extended Data Fig. 9: Human WI-38 fibroblast senescence validation.The alternative text for this image may have been generated using AI.

a, Phase contrast micrographs of SA-βgal activity (blue) staining in human WI-38 fibroblasts that were either proliferating (P), rendered senescent by replicative senescence (RS), or by treatment with Doxo, IR and additional culture for 7–10 days. b, RT-qPCR analysis of the levels of CDKN2A, CDKN1A, GDF15, LMNB1 and IL8 mRNAs in WI-38 fibroblasts treated as described in (a). c-f, RT-qPCR analysis of the levels of SPP1, SERPINE2, CTSB, TNFRSF11B, PRG4, LTPBP2, FTH1, MGP, LUM, CD9, APOE, LGALS3, CST3, SPARC, THBS1, MMP3, CTSS, and LCP1 mRNAs in human WI-38 fibroblasts treated as described in (a). In b-f, data represent the means ± SD for P, n = 3; IR, n = 3; n = 3; Doxo, n = 3; RS, n = 3; n represents biological replicates. In panel (b), CDKN2A, IR (p = 0.0099), Doxo (p ≤ 0.0001), RS (p = 0.0005) vs. P; CDKN1A, IR (p ≤ 0.0001), Doxo (p ≤ 0.0001), RS (p ≤ 0.0001) vs. P; GDF15, IR (p = 0.001), Doxo (p ≤ 0.0001), RS (p ≤ 0.0021) vs. P; LMNB1, IR (p = 0.001), Doxo (p = 0.0002), RS (p = 0.0009) vs. P; IL8, IR (p ≤ 0.0001), Doxo (p = 0.0005), RS (p = 0.0003) vs. P. In panel (c), SPP1, IR (p = 0.0085), Doxo (p = 0.0035), RS (p = 0.0071) vs. P; SERPINE2, IR (p = 0.0014), Doxo (p = 0.0025), RS (p = 0.0026) vs. P; CTSB, IR (p = 0.0003), Doxo (p = 0.0002), RS (p = 0.0007) vs. P; TNFRSF11B, IR (p = 0.0013), Doxo (p = 0.0003), RS (p ≤ 0.0001) vs. P; PRG4, IR (p = 0.0135), Doxo (p = 0.0043), RS (p = 0.0039) vs. P. In panel (d), LTPBP2, IR (p = 0.0003), Doxo (p = 0.0012), RS (p = 0.0006) vs. P; FTH1, IR (p = 0.0025), Doxo (p = 0.0038), RS (p = 0.0016) vs. P; MGP, IR (p = 0.0007), Doxo (p ≤ 0.0001), RS (p ≤ 0.0001) vs. P; LUM, IR (p = 0.0005), Doxo (p = 0.0059), RS (p = 0.0014) vs. P; CD9, IR (p = 0.0051), Doxo (p = 0.7688), RS (p = 0.0008) vs. P. In panel (e), APOE, IR (p = 0.0008), Doxo (p = 0.007), RS (p ≤ 0.0001) vs. P; LGALS3, IR (p = 0.0108), Doxo (p = 0.0229), RS (p ≤ 0.0001) vs. P; CST3, IR (p = 0.0026), Doxo (p ≤ 0.0001), RS (p ≤ 0.0001) vs. P; SPARC, IR (p = 0.0021), Doxo (p = 0.002), RS (p = 0.0004) vs. P; THBS1, IR (p = 0.0003), Doxo (p = 0.0114), RS (p ≤ 0.0001) vs. P. In panel (f), MMP3, IR (p = 0.0015), Doxo (p = 0.0007), RS (p = 0.0003) vs. P; CTSS, IR (p = 0.0002), Doxo (p = 0.0023), RS (p = 0.0019) vs. P; LCP1, IR (p = 0.0003), Doxo (p = 0.9169), RS (p ≤ 0.0001) vs. P.Significance was established using Shapiro-Wilk test first, followed by Welch’s t-test. *, p ≤ 0.05; **, p ≤ 0.01; ***, p ≤ 0.001; ****, p ≤ 0.0001.

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