Extended Data Fig. 1: Telomerase activity, cell death, and fibroblast line validation.
From: Evidence for improved DNA repair in the long-lived bowhead whale
a, Telomerase activity in bowhead whale tissues. MSF, mouse skin fibroblasts, and HeLa cells used as a positive control. b, qRT–PCR analysis of hTERT expression in primary and hTERT-immortalized fibroblasts from human and bowhead whale (n = 3 biological replicates per species). Data are presented as mean ± SD. c, Apoptosis and necrosis in fibroblasts from human (n = 3 biological replicates) and bowhead whale (n = 2 biological replicates) following ENU treatment (3 h) at indicated doses. Annexin V/PI staining was performed after 3 days. Data are presented as mean. No statistical tests were applied because n < 3 for bowhead whale samples. d–j, Western blots validating fibroblast cell lines. d, SV40 Large T antigen (LT) and H-RasG12V in transformed vs. untransformed lines (Fig. 2). e, SV40 Small T antigen (ST) in the same lines. f, p53 in colonies after CRISPR-mediated TP53 knockout; clones C13 and C17 were selected. g, Rb in colonies after CRISPR-mediated RB1 knockout using pooled p53–/– clones; clones C1 and C26 were selected. (h) p53 in clones C13 and C17. i, Rb in clones C1 and C26. j, PTEN in colonies after CRISPR-mediated PTEN knockout. For gel source data, see Supplementary Figure 1. k, l, Reporter assays confirming knockout clones. k, p53 activity measured by firefly/Renilla luciferase after etoposide treatment. l, Rb activity measured by E2F-responsive firefly/Renilla luciferase; elevated E2F reflects reduced Rb function. n = 3 biologically independent experiments. Data are presented as mean ± SD; P values were calculated using Welch’s two-sided t-test.
