Fig. 2: Acetate-mediated histone acetylation and ACSS2 activation control pancreatic cancer growth under acidosis.
a, Relative expression of SREBF1, ACACA and FASN genes in PDAC cells treated with 5 mM acetate under acidosis (n = 4 in each group, from independent biological replicates). b,c, Representative fluorescent images (b) and quantification (c) of Nile red staining in PDAC cells treated with 5 and 10 mM acetate. Scale bars, 100 µm (n = 5 in each group, from independent biological replicates). d, Relative survival of PDAC cells treated with vehicle or 100 µM orlistat, without or with acetate (n = 5 in each group, from independent biological replicates). e, Immunoblots of acetylated histones in PDAC cells treated with acetate (SE, short exposure; LE, long exposure). Images are representative of two independent experiments. f, Immunoblots of acetylated histones in PDAC cells treated with CM from shScr, shACLY-a and shACLY-b HPS cells. Images are representative of two independent experiments. g, 14C-acetate incorporation (counts per minute (c.p.m.)) in total histones extracted from S2-013 and HPAF-II cells treated with acetate in the absence and presence of 10 µM C646 for 6 h under acidosis (n = 3 in each group, from independent biological replicates). h, Relative survival of PDAC cells treated with 10 µM C646 inhibitor, without or with acetate (n = 5 in each group, from independent biological replicates). i, Relative mRNA expression of ACSS2 in PDAC cells treated with acetate under acidosis (n = 4 in each group, from independent biological replicates). j, Immunoblot of ACSS2 in PDAC cells upon treatment with acetate for 24 h under acidosis. Representative of two independent experiments. k,l, Representative immunofluorescent images (k) and quantitation (l) showing expression of ACSS2 protein in acidic tumour regions, as imaged by staining with pHLIP in tumour sections from 20-week-old KPC mice, and AT-nude mice orthotopically implanted with S2-013 cancer cells. Scale bars, 100 µm (KPC, normal (n = 24) and acidic (n = 24) pH; S2-013, normal (n = 21) and acidic (n = 168) pH). Unpaired two-tailed t-test, mean ± s.e.m. (a,g,i,l); one-way ANOVA with Bonferroni’s post-hoc test, mean ± s.e.m. (c); two-way ANOVA with Tukey’s post-hoc test, mean ± s.e.m. (d,h).
