Skip to main content
. Author manuscript; available in PMC: 2023 Nov 20.
Published in final edited form as: Cancer Res. 2023 May 15;83(10):1596–1610. doi: 10.1158/0008-5472.CAN-22-2316

Figure 1: A hypoxic signature is enriched in inflammatory fibroblasts in PDAC.

Figure 1:

(A) Single sample Gene Set Enrichment Analysis (ssGSEA) of selected hallmark signatures in myofibroblastic CAFs (myCAFs) and inflammatory CAFs (iCAFs) based on single-cell RNA-sequencing (scRNA-seq) data from human PDAC. Data from (5).

(B) Schematic of experimental workflow to analyze Pimonidazole enrichment and localization in mouse PDAC tumors arising from orthotopic transplantation of KPC organoids.

(C-E) Analysis of Pimonidazole in Ly6C+ and Ly6C− cells among live, CD31−CD45−EpCAM−PDPN+ cells in PDAC tumors. (C) Gating for Ly6C in PDPN+ cells. (D) Histogram of fluorescence intensity and (E) quantification of Pimonidazole median fluorescence intensity (MFI) comparing Ly6C+ and Ly6C− cells. A.U. = arbitrary units. N=3 mice. P-value was calculated by ratio paired t-test.

(F, G) Immunofluorescence staining of Pimonidazole, PDPN and αSMA in mouse PDAC tumors. (F) Representative image. Nuclei are labeled with DAPI. Scale bar = 500 μm. (G) Quantification of αSMA− and αSMA+ pixel among PDPN+ pixel within Pimonidazole-stained regions. N=8 sections from 4 mice. Data represent mean+SD. P-value was calculated by ratio paired t-test.

(H) GSEA comparing PSCs cultured in normoxia (20% O2) or hypoxia (0.5% O2) for 48h. iCAF signature derived from (4). Other signatures represent Hallmark signatures from MSigDB. N=3 biological replicates.