Fig 2: GCV treatment of p16-3MR mice does not affect atherosclerosis, but induces inflammation. (A) Aortic root plaques in ApoE→ApoE, p16→ApoE, ApoE→p16, or p16→p16 mice + GCV, or p16→p16 mice + saline, stained with Masson’s trichrome, TUNEL, or Mac3. Scale bar = 300 µm. High power inset shows apoptotic cell and nuclear debris from outlined area. (B) Plaque area for mice in (A). (C and D) Number of TUNEL+ cells/aortic root plaque (C) or %Mac3+ cells (D) for mice in (A). (E–G) Relative mRNA expression for p16, IL18, or TNFα in experimental mice. Data are means (SD) n = 5–10 mice. One-way ANOVA with correction for multiple comparisons (B–D) or Kruskal–Wallis H test followed by Dunn’s multiple comparisons test (E–G).

Fig 3: ABT-263 reduces atherosclerosis, but not local SASP cytokine expression. (A) ORO staining of mouse descending aorta treated with control (vehicle) or ABT-263, and quantification of %ORO area (n = 11–14). Scale bar = 3 mm. (B) Masson’s trichrome histochemistry of aortic root atherosclerotic plaque from mice treated in (A). Panels below show high power view of outlined area. Arrow shows necrotic core. Scale bar = 200 µm. (C–E) Aortic root plaque area/total area (C) Cap area (D), or Core area (E) for mice in (A). n = 11–13. (F) qPCR for relative expression of p16 or SASP cytokines in aortic arches of experimental mice against the housekeeping gene HMBS (n = 7). Data are means (SD), n = 10. Unpaired Student’s t-test (A, C–E) or Mann–Whitney U test (F).

Fig 4: ABT-263 (Navitoclax) selectively reduces senescent VSMCs. (A and B) Photomicrographs (A) or quantification (B) of mouse VSMCs stained for SAβG, as replicating control cells or after dox1 + 7 days treatment, or each group ±1 µM ABT-263 treatment for 48 h. (C) Western blot and quantification for p16 in cells treated in (A and B). (D) Fold change in mRNA expression compared with control replicating cells for p16 and a range of SASP cytokines against the housekeeping gene HMBS. Data are means (SD), n = 4–5. Unpaired Student’s t-test. (E) Mouse macrophages cultured for 28 days, then treated with 1 µM ABT-263 for 48 h and stained for SAβG. Data are means (SD), n = 3. Unpaired Student’s t-test.

Fig 5: Correlation of CNV of CCND1, CDKN2A and RAD52 with high EBV copy number in 22 NPC plasma. (a) CNV of CCND1, CDKN2A and RAD52 in 22 NPC plasma with high EBV DNA load (> 5000 copies/ml) based on Q-PCR results. (b) Correlation plot between CNV of CCND1, CDKN2A and RAD52 and log EBV DNA load in 22 NPC plasma samples (c) Correlation between CNV of the CCND1/CDKN2A ratio and log EBV load in 22 NPC plasma. Pearson’s correlation coefficient, r, and equations of regression are indicated