Fig. 5: SAT1 expression in PDAC cells is critical for acetate-mediated growth of pancreatic cancer cells in vitro and in vivo.
a,b, Relative levels of SAT1 mRNA and protein in control (shScr) and SAT1 knockdown (shSAT1-a and shSAT1-b) S2-013 (a) and HPAF-II (b) cells under acidosis (n = 4 in each group, from independent biological replicates). Images are representative of two independent experiments. c,d, Relative survival of shScr and SAT1 knockdown S2-013 (c) and HPAF-II (d) cells in the presence and absence of acetate under acidosis (n = 3 in each group, from independent biological replicates). e,f, Relative levels of polyamine biosynthetic pathway metabolites in shScr and shSAT1 S2-013 (e) and HPAF-II (f) cells in the presence and absence of 5 mM acetate under acidosis. Metabolites are presented as normalized row Z-scores from n = 6 (S2-013) and n = 4 (HPAF-II) biological replicates per group. g,h, Representative images (g) of PA417 and PA901 organoids cultured alone or in combination with HPS cells upon treatment with 25 µM pentamidine, along with mean organoid diameters (h). Scale bars, 100 µm. PA417, n = 4 (vehicle), n = 4 (+HPS), n = 6 (pentamidine), n = 4 (HPS + pentamidine), and PA901, n = 4 in each group from independent biological replicates. i,j, Tumour weights (i) and volumes (j) upon necropsy of mice implanted with control and SAT1 knockdown S2-013 cells alone or co-implanted with HPS cells (n = 12 (shScr), n = 12 (shScr + HPS), n = 10 (shSAT1), n = 10 (shSAT1 + HPS) mice). k,l, Representative IHC images for Ki-67 staining (k) in tumour sections from mice implanted with shScr or shSAT1 S2-013 cells alone or co-implanted with HPS cells along with the quantitation of percent positive cells (l). Scale bars, 100 µm. Ki-67 positive cells were counted in three different fields from three tumour sections of each group (n = 9 in each group). One-way ANOVA with Bonferroni’s post-hoc test, mean ± s.e.m. (a,b,h–j,l); two-way ANOVA with Tukey’s post-hoc test, mean ± s.e.m. (c,d).
