Fig 2: Single-cell RNA-seq identifies a combination of fibrocyte specific markers.A Schematic diagram of CD45+ cell isolation from the bone marrow of a C57BL/6 mouse. B Uniform manifold approximation and projection (UMAP) representation of the data clustered by the Leiden algorithm revealing 17 clusters (left). Expression of Prprc (CD45) is shown in the panel on the right. C UMAP representation of all cells belonging to cluster 13, 14, 16, and 17 by the Leiden algorithm. Expression of Serpinh1, Ccr5, Cxcr3, Selplg, Cd44, Ccr2, Cd163, S100a8 and Cd9 are shown. D Representative FACS contour plots of fibrocytes (CD45+SiglecH–, CD162+CD9+CD11b+CD44+CCR2/5+) in bone marrow of C57BL/6 mice. (E) Representative FACS contour plot and histogram showing Ly6C, F4/80, and Col1 expression in monocytes (CD11b+/Ly6G-; orange), lymphocytes (CD11b–; blue) and fibrocytes (CD45+/SiglecH–, CD162+CD9+CD11b+CD44+CCR2/5+; red). F Quantification of fibrocytes in fibrocyte-depleted mice (HSV-TK/Col1+Ganciclovir) and control mice (HSV-TK/Col1), n = 4. p-values were determined by two-tailed unpaired t–test with Welch’s correction. G Representative FACS contour plots of fibrocytes in fibrocyte-depleted (HSV-TK/Col1+Ganciclovir) and control (HSV-TK/Col1) mice. Source data are provided in the source data file.

Fig 3: Lung cancer CD45+ cells scRNA-seq validates identified fibrocyte specific markers.A Schematic diagram of CD45+ cell isolation from the healthy, LLC1 and KRasLA2 lung tumor models. B UMAP representation of the data clustered by the Leiden algorithm revealing 15 clusters (left). Expression of Prprc (CD45) is shown in the panel on the right. C UMAP representation of all cells belonging to cluster 12, 13, and 14 by the Leiden algorithm. Expression of Serpinh1, Ccr5, Cxcr3, Selplg, Cd44, Ccr2, Cd9, Cd68, and Csf1r are shown. D Gating strategy used to identify fibrocytes, monocytes, macrophages, neutrophils and osteoblast-like cells. Representative flow-cytometric analysis used to identify fibrocytes (CD45+CD11b+CD162+F4/80+CCR2/5+CD9+CD44+Col1+), monocytes (CD45+CD11b+/Ly6C+), macrophages (CD45+CD11b+CD162+F4/80+CCR2/5lo/–CD9–), neutrophils (CD45+CD11b+Ly6G+) and osteoblast-like cells (CD45–CD44+CD9+ RANKL+). E Representative FACS histograms show expression of Col-1, CD44, CXCR3 in fibrocytes (CD45+CD162+F4/80+CD9+CCR2/5+), macrophages (CD45+CD162+F4/80+CD9–CCR2/5–), monocytes (CD45+CD162+Ly6C+), neutrophils (CD45+Ly6G+) and osteoblast-like cells (CD45–CD9+CD44+RANKL+). F Quantification of osteoblast-like cells (CD45–CD9+CD44+RANKL+) in the i.v. lung tumor model from fibrocyte-depleted (HSV-TK/Col1+Ganciclovir), HSV-TK/Col1 compared with control lung tumor tissue by FACS analysis. n = 5. p-values were determined by One-way ANOVA with Fisher’s LSD test. Source data are provided in the source data file.