Figure 3.

S100A8 polarizes CAFs toward myCAFs

(A) Spearman correlations between the S100A8 RNA level of epithelial cells and the myCAF (left) and iCAF (right) signature scores of fibroblasts. The gray areas represent 95% CIs (n = 60).

(B) Representative multiple immunofluorescence images of KRT6A, S100A8, and αSMA in the PDX donors’ primary tumor tissues of the R (n = 8) and NR (n = 12) groups. Scale bar, 100 μm.

(C) Quantification of S100A8 and αSMA fluorescence intensity in (B). Data are presented as mean ± SEM. p values are determined using two-tailed Student’s t test.

(D) Spearman correlation between the S100A8 and the αSMA fluorescence intensity. The gray area represents 95% CI (n = 20).

(E) Western blot analysis of the myCAF-related marker proteins collagen type 1 and αSMA, the iCAF-related marker CXCL1 in CAFs cocultured with KYSE510 and KYSE180 cells with or without S100A8 knockdown.

(F) Representative images of CAF migration induced by CM derived of KYSE510 and KYSE180 cells with or without S100A8 knockdown. Scale bar, 100 μm.

(G) Quantification of (F) (n = 3 biological replicates). Data are presented as mean ± SD. p values are determined using two-tailed Student’s t test.

(H) Western blot analysis of collagen type 1, αSMA, and CXCL1 in CAFs treated with indicated concentration of rS100A8 proteins.

(I) Representative images of CAF migration induced by indicated concentration of rS100A8 proteins. Scale bar, 100 μm.

(J) Quantification of (I) (n = 3 biological replicates). Data are presented as mean ± SD. p value is determined using two-tailed Student’s t test.

For all panels, ∗∗p < 0.01, ∗∗∗p < 0.001. Each assay for western blot had three biological repeats. See also Figure S3.