Extended Data Fig. 9: Preclinical profiling of ROS generation, liver physiological index, genome-wide expression and physical function of aged mice.
From: PDK4-dependent hypercatabolism and lactate production of senescent cells promotes cancer malignancy
a. Immunoblot analysis of NOX1 expression across organ types in young vs aged mice. Y, young. A, aged. Myo, myocardium. GAPDH, loading control. b. Measurement of ROS levels with DCFH-DA. Left, representative images. Right, statistical comparison. Scale bar, 20 μm. c. Heat map depicting expression pattern of genes significantly upregulated upon BLEO-induced senescence but subject to counteraction by PDK4-IN. Red stars, representative SASP factors. d. GSEA plot of a significant gene set in SASP spectrum. FDR, false discovery rate; NES, normalized enrichment score. e. The serum levels of alanine transaminase (ALT) from young mice and their aged counterparts that received biweekly treatment as indicated at 20 months of age for 4 consecutive months. f. The serum levels of aspartate transaminase (AST). g. The serum levels of lactate dehydrogenase (LDH). h. Measurement of body weights. i. Assessment of daily food intake. j. Whole-life survival curves of C57BL/6 J mice treated biweekly with vehicle (n = 68; 37 males, 31 females), PDK4-IN (n = 66; 33 males, 33 females) or PCC1 (n = 67; 35 males, 32 females) starting at 24-27 months of age. k-l. Maximal walking speed (k) and hanging endurance (l) averaged over the last 2 months of life. m. Appraisal of lifespan for the longest living mice (top 20) per group. Animals were adapted in 3 (young) or 4 (aged) independent cages. For in vitro experiments, n = 3 for b. For preclinical assays, n = 10/group for e-i and k-m. Data in b, e-i and k-m are shown as mean ± S.D. For box and whiskers graphs (h-i, k-l), the minima, maxima, median, 25th and 75th percentiles are shown, with whiskers indicating smallest and largest values. P values were calculated by two-sided unpaired Student’s t-tests (b, e, f, g, m), one-way ANOVA (h-i, k-l) or Log-rank (Mantel–Cox) tests (j). For data in d, P value was calculated by one-way ANOVA with Tukey’s post hoc comparison. ^, P > 0.05. *, P < 0.05. **, P < 0.01. ***, P < 0.001. ****, P < 0.0001.
