Extended Data Figure 1. ATTAC transgene expression tracks with expression of senescence markers in iWAT and induces apoptosis of senescent cells upon AP administration.

a, Comparative analysis of SA-β-Gal activity in intact iWAT. b, Analysis of endogenous p16^Ink4a and ATTAC transcript SA-β-Gal activity in iWAT by qRT-PCR. Abbreviation: H/H (BubR1^H/H) (n = 4 mice per group). Scale bar, 0.5 cm. c, FACS-based quantitation of iWAT progenitor cell numbers in 18-month-old ATTAC mice treated with vehicle or AP. d, Expression of the ATTAC transgene and senescence markers in iWAT as determined by qRT-PCR (n = 4 mice per group). Asterisks above individual bars denote significant changes to 2-month-old mice; asterisks above brackets denote significant differences between 18-month-old vehicle and AP treated mice. e, Perirenal, mesenteric, subscapular and brown adipose depot weights. f, SA-β-Gal activity in iWAT from 2-month-old ATTAC mice treated with vehicle or AP beginning at weaning age. g, p16^Ink4a levels in iWAT from the mice described in f. Actin was used a loading control. h, Expression of ATTAC and senescence marker mRNA in the mice described in f (n = 3 mice per group). (i–k) Early passage non-senescent ATTAC MEFs express p16^Ink4a but are not susceptible to FKBP-Casp8-mediated elimination when cultured in the presence of AP. i, Levels of p16^Ink4a in passage 3 ATTAC mouse embryonic fibroblasts (MEFs), with and without AP treatment. j, Growth curves of passage 3 ATTAC MEFs (n =4 independently generated MEF lines per group), with and without AP treatment. k, Expression of ATTAC and senescence marker mRNA in passage 3 ATTAC MEFs (n = 3 independently generated MEF lines per group), with and without AP treatment. Error bars indicate s.e.m. Statistical significance was determined by unpaired two-tailed t tests. *, P<0.05; **, P<0.01; ***, P<0.001. For gel source data, see Supplementary Fig. 1.