Fig. 2. Senescent CAFs are present in human pancreatic cancer lesions.

a Numbers of human premalignant lesions and PDACs in which p16+ cells were detected in the tumor or stromal compartments, in both, or in neither, as indicated. P value, Chi-squared, refers to the difference in distribution between the lesion types. b Sections of human premalignant (low grade PanIN) (left) and PDAC (right) samples, stained for p16, as scored in panel a. Black arrows indicate p16+ CAFs. c Co-stain of human PDAC sections for p16, the epithelial marker CK18, and vimentin (VIM), as scored in (a). Arrows indicate p16+ CAFs. d FACS isolation of SA-βGal- and SA-βGal+ CAFs from a representative human PDAC sample. CD90 marks fibroblasts. Plot on right is gated for EpCAM-CD45- cells. Gate values are calculated out of CD90+ fraction. e Average percentage of SA-βGal+ cells in CAFs from different patients. Mean of n = 5 tumors ± SEM. f Levels of p16 (CDKN2A) mRNA in matched SA-βGal- and SA-βGal+ CAFs isolated from human PDAC patients. Mean of n = 4 tumors ± SEM. t test. g UMAPs showing a scRNA-seq cluster of 3492 individual PDAC CAFs from 21 patients, obtained from ref. ³¹. Red color indicates the relative scores of iCAF, myCAF and apCAF expression signatures in individual CAFs. h UMAP of same CAFs, indicating relative expression score of the senescence signature. i Percentage of CAFs scoring positive for the senescence signature in each of the individual subjects included in the scRNA-Seq analysis. Mean of n = 21 patients ± SEM. j Percentages of CAFs classified to the indicated subtypes which score positive for the senescence signature. Scale bars = 20 μm.