Extended Data Fig. 3: Characterizing acetate secretion in different CAF subtypes.
a,b Representative images showing expression of αSMA (a) or Vimentin (b) by immunofluorescence staining in HPS and CAF-0911 cell lines and primary CAFs (CAF-0906, CAF-1003, and CAF-1016), as imaged by staining cells with fluorescently-tagged phalloidin for F-actin. Nuclei were stained with DAPI. Scale bar = 20 µm. c, Relative mRNA expression of iCAF markers (IL6, LIF) and myCAF markers (ACTA2, MYL1) in Huff1 (human foreskin fibroblast cell line; used as a control), CAF-0911, CAF-0906, CAF-1003, and CAF-1016 cells. The gene expression is normalized to Huff1 cells (n = 4 in each group from independent biological replicates). d, Acetate levels in Huff1 control cells and CAF-0911, CAF-0906, CAF-1003, and CAF-1016 cancer-associated fibroblast cells (n = 3 in each group from independent biological replicates). e,f, Acetate levels in HPS and CAF-0911 cells upon treatment with IL-1β (e) or Tgfβ1 (f) for 48 hrs (n = 3 in each group from independent biological replicates). g,h, Acetate estimation in CAF-0906 and CAF-1003 cells upon treatment with IL-1β (g) or Tgfβ1 (h) for 48 hrs. (n = 3 in each group from independent biological replicates). One-way ANOVA with Bonferroni’s post-hoc test; mean ± s.e.m. (c,d); unpaired, two-tailed t-test; mean ± s.e.m. (e,f,g,h).
