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. 2022 Jan 28;13:100101. doi: 10.1016/j.mbplus.2022.100101

Fig. 2.

Fig. 2

CAFs and MR+ TAMs are the main cell types in collagen internalization. (A-C) Quantification of collagen internalization by cell types of the tumor microenvironment: Single-cell suspensions of LL/2 tumors (A), LL/2-GFP tumors (B) or EO771.LMB-GFP tumors (C) were cultured overnight with soluble 10 µg/mL A647-labeled collagen type I. Flow cytometry analysis was then performed to determine the internalized fluorescence in CAFs, TAMs, dendritic cells, granulocytes, monocytes, cancer cells, CD45−CD31−FAP− cells and CD45−CD31−FAP−GFP− cells. n = 2–6. Error bars = SEM. (D) Representative density plots from flow cytometry analysis of LL/2 tumor showing how TAMs were divided into subpopulations negative and positive for MR. Gates were based on isotype control. (E) Representative overlay of histograms from flowcytometry analysis of LL/2 tumors showing the level of internalized collagen in TAMs negative and positive for MR. (F-G) Quantification of collagen internalization by MR+ macrophages, MR− macrophages and FAP+ fibroblasts from LL/2 (F) and EO771.LMB tumors (G). n = 3–5, except CAFs in 3F (n = 2). Error bars = SEM. *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001; one-way ANOVA with post hoc Tukey’s multiple comparisons test.