Fig. 2. Senescent cell’s partial removal increases the survival of GBM-bearing mice.
a Timeline of tumorigenesis induction (lv: H-RasV12-GFP-shp53) and removal of senescent cells with GCV, 21 days post lv injection in the p16-3MR transgenic mouse. b Timeline of tumorigenesis induction (lv-luc: H-RasV12-GFP-P2A-Luc2-shp53) and removal of senescent cells with GCV in the p16-3MR mouse or with ABT263 in WT mouse when head to body bioluminescence ratio reached 2. c Kaplan–Meier survival curves (solid lines) of WT (n = 10, median survival 38 days) and p16-3MR (n = 9, median survival 51 days) mice treated with GCV as shown in (a). Kaplan–Meier survival curves (dotted lines) of p16-3MR mice treated with vhc (n = 14, median survival 36 days) or GCV (n = 15, median survival 46 days) as shown in (b). d Kaplan–Meier survival curves of WT mice treated with vhc (n = 11, median survival 34 days) or ABT263 (n = 11, median survival 46 days) as shown in (b). e Representative HE, GFP IHC, and SA-β-Gal staining on adjacent mouse GBM cryosections. Right panels represent higher magnifications of the left panels. Scale bars, left panels: 2.5 mm, right panels: 20 µm. f Quantification of the SA-β-gal area over the tumor (GFP+) area (n = 7 biologically independent animals/group). g Relative transcript levels of p16Ink4a, shown as FPKM estimates extracted from bulk RNAseq analysis (WT+GCV, n = 5; p16-3MR+GCV, n = 9). h Relative transcript levels are shown as ratios of normalized values of p16-3MR+GCV GBMs (n = 6) over WT+GCV GBMs (n = 4). i GSEA graphs from bulk RNAseq data in p16-3MR+GCV GBMs compared with WT+GCV GBMs. SASP gene list from Gorgoulis et al.3 (Supplementary Data 1). j Relative transcript levels of genes in WT+GCV and p16-3MR+GCV GBMs extracted from bulk RNAseq data (WT+GCV, n = 5; p16-3MR+GCV, n = 9). c and d Statistical significance was determined by Mantel–Cox log-rank test. b Raw p-values from the log-rank tests are included in the figure and significance is indicated by * for p-values below the 5% level after correction by the Benjamini–Hochberg procedure. f–h, j Data are represented as the mean ± SD and statistical significance was determined by the Wilcoxon–Mann–Whitney test (*p < 0.05; **p < 0.01, ns, not significant). TMX tamoxifen, vhc vehicle, gav. gavage, GCV ganciclovir, i.p. intraperitoneal, lv lentivirus, lv-luc lentivirus-luciferase, GSEA gene set enrichment analysis, FDR false discovery rate, NES normalized enrichment score, r. enrichment score running enrichment score. Raw data are provided as a Source Data file.