Extended Data Fig. 1: Additional characterization of pancreatic stellate cell-mediated support of cancer cell growth by secretion of acetate.
a, Relative cell survival of pancreatic cancer cells (S2-013, HPAF-II, CFPAC-1, T3M4, and MIAPaCa-2) upon treatment with stellate cell (CAF-0911 and HPS)-derived conditioned medium (CM) (n = 4 for S2-013, n = 4 for HPAF-II, n = 6 for CFPAC-1, n = 4 for T3M4, n = 4 for MIAPaCa-2 in each group from independent biological replicates). b,c, Representative images and quantitation of diameter of PA417 (b) and PA901 (c) organoids cultured in the presence or absence of HPS cells. Scale bar = 100 µm (PA417), 250 µm (PA901) (n = 6 in each group from independent biological replicates). d, Immunofluorescent images showing the distribution of HPS cells stably expressing LeGO-dKatushka2 plasmid when co-cultured with multiple pancreatic cancer organoids (PA417, PA901, and PA137) labeled with CellTrace Violet dye for 5 days. Scale bar = 100 µm. Representative image of two independent experiments. e, Relative fold change in labeled metabolite levels in CM from CAF-0911 stellate cells (CM) or double conditioned medium (DCM) derived from CAF-0911 cells pre-conditioned with S2-013 tumor cell-conditioned media (n = 3 in each group from independent biological replicates). f, Levels of acetate in the interstitial fluid of pancreatic cancer tissues of KPC mice (female mice at 20-22 weeks of age) and control C57BL/6 J mice (n = 5 in each group from independent biological replicates). g-i, Relative cell survival of pancreatic cancer cells, S2-013 (g), HPAF-II (h), and CFPAC-1 (i) treated with increasing doses of acetate (0.1–10 mM) for 72 hrs cultured in normoxic, hypoxic, low glucose, and low glutamine conditions. The cell survival is normalized to the respective untreated controls (n = 6 in each group from independent biological replicates). One-way ANOVA with Tukey’s post-hoc test; mean ± s.e.m. (a,g,h,i); unpaired, two-tailed t-test; mean ± s.e.m. (b,c,e,f).
