Fig. 2: PDAC patients with poor prognosis have elevated levels of intratumoral collagen, PlGF/VEGF-A, and CAFs expressing NRP1/VEGFR1/VEGFR2/PD-L1.

Correlation analyses between a tumor fibrosis (measured by Masson’s trichrome⁺ area) and overall survival of PDAC patients, between PlGF⁺ area% (detected by IHC staining) and b overall survival or c disease-free survival of PDAC patients, between d VEGFA⁺ area% (detected by IHC staining) and overall survival of PDAC patients, and between tumor fibrosis and e PlGF⁺ area% or f VEGFA⁺ area%. Data from 20 PDAC patient samples were analyzed (n = 20). Statistical significance was accessed by two-tailed Student’s t test. g Double IHC staining for α-SMA with NRP1, VEGFR1, or VEGFR2 in tumors from PDAC patients with poor prognosis and good prognosis. Images were produced via Image J. Red color and DAB color were separated via color deconvolution of IHC images and merged using color merge functions. The red color represents α-SMA in CAFs, the green color represents each indicated receptor, and the yellow color (merged area) indicates co-expression of the two proteins. Four random spots additionally examined showed similar results. Scale bar, 100 μm. h–k Correlation analyses between overall survival of PDAC patients and h NRP1⁺α-SMA⁺ area%, i VR1 (VEGFR1)⁺α-SMA⁺ area%, or j VR2 (VEGFR2)⁺α-SMA⁺ area%, and between k tumor fibrosis and NRP1⁺α-SMA⁺ area%. Data from 20 PDAC patient samples were analyzed (n = 20). Statistical significance was accessed by two-tailed Student’s t test. l IHC staining for α-SMA and PD-L1. Red, α-SMA in CAFs; green, PD-L1; yellow, co-expression of α-SMA and PD-L1. Four random spots additionally examined showed similar results. Correlation analyses between PD-L1⁺α-SMA⁺ area% and m overall survival of PDAC patients or n tumor fibrosis. Data from 20 PDAC patient samples were analyzed (n = 20). Statistical significance was accessed by two-tailed Student’s t test. o Flow cytometry analysis of the ratio of PD-L1⁺ cells (indicated) over Pan02 tumor-infiltrating live cells. Data from six Pan02 tumor-bearing mouse-derived tumor samples were analyzed (n = 6). Data are presented as the mean ± SEM. Cancer, cancer cell; E.C., endothelial cell. Source data are provided as a Source Data file.