Fig. 1

Identification of four CAF subsets in HGSOC. a Representative view of HES staining of non-mesenchymal and mesenchymal HGSOC sections (Institut Curie cohort). Scale bar, 100 μm (low magnification) and 40 μm (inset). b Scatter plot showing the percentage of stroma in HGSOC. N = 107. Data are shown as mean ± SEM. P values are from Mann-Whitney test. c Bar plot showing association of mesenchymal HGSOC with stromal features, defined by pathologists as loose (low stromal cellularity) or dense (high stromal cellularity). N = 56. P value is from Fisher’s Exact Test. d Gating strategy to identify CAF subsets in HGSOC by FACS. Results from a representative HGSOC patient are shown. Cells isolated from freshly dissociated human HGSOC were first gated on DAPI⁻, EPCAM⁻, CD45⁻, CD31⁻ cells, for excluding dead cells (DAPI⁺), epithelial cells (EPCAM⁺), hematopoietic cells (CD45⁺), and endothelial cells (CD31⁺). Selected cells were next examined using six fibroblast markers. Representative flow cytometry plots show gating strategies based on FAP, CD29, SMA, and FSP1 that allow the identification of four sub-populations of fibroblasts in HGSOC: CAF-S1 are CD29^Med FAP^High SMA^High FSP1^High, CAF-S4 are CD29^High FAP^Low SMA^High FSP1^Med, CAF-S3 are CD29^Low FAP^Low SMA^Low FSP1^Med/High and CAF-S2 are CD29^Low FAP^Low SMA^Low FSP1^Low. e CytoSpade trees annotated with each marker expression in HGSOC analyzed by FACS. Colors show staining intensity for each marker. Size of the nodes is proportional to the number of cells showing similar staining for the markers analyzed. f Scatter plots showing specific mean fluorescent intensity (MFI) detected for each marker in each CAF sub-population. Each dot represents the specific median of fluorescent intensity of the cellular population by patient. N = 22. Data are shown as mean ± SEM. P values are from Student’s t-test