Fig. 7: CAF-S1 and CAF-S4 promote cancer cell invasion by TGFβ, CXCL12 and NOTCH. | Nature Communications

Fig. 7: CAF-S1 and CAF-S4 promote cancer cell invasion by TGFβ, CXCL12 and NOTCH.

From: Cancer-associated fibroblast heterogeneity in axillary lymph nodes drives metastases in breast cancer through complementary mechanisms

Fig. 7

a Up, Representative images of E-cadherin (red), F-actin (green) and DAPI (blue) in BC cells alone (−) or with CAF-S1 transfected with non-targeting (siCTL) or CXCL12-targeting (siCXCL12) siRNA. CAF-S4 images in Supplementary Fig. 5c. Scale bars, 20 μm; inset 10 μm. Down, BC cell density and E-cadherin staining alone (gray) or with CAF (CAF-S1 red; CAF-S4 blue) (≥12 images/condition; n = 8). b Same as a with siCTL- or siCXCR4-transfected BC cells, with/without CAF-S1 (red/gray) (≥8 images/condition; n = 4). c Same as a with/without CAF-S1 with/without TGFβ-R inhibitor (LY2109761). CAF-S4 images in Supplementary Fig. 5e. Quantifications without (gray) or with CAF (CAF-S1 red; CAF-S4 blue) (≥7 images/condition; MCF7: n = 7; T47D: n = 3). d Percentage (%) of collagen contraction by CAF-S4 (blue) or CAF-S1 (red) without (DMSO) or with DAPT (n ≥ 3). e Strain energy density of CAF-S4 without (DMSO, blue) or with DAPT (gray). Each dot is one cell, n ≥ 46 cells/condition. f Representative 3D views of MDA-MB-231 (green) invasion by inverted Transwell assays in CAF-S4 (blue)-embedded collagen matrix without (DMSO) or with DAPT. Colors indicate distance browsed by BC cells on z axis (dmax180 μm, red). Scale bar, 50 μm. g Maximal distance of BC cells in CAF-S4-embedded collagen with DAPT relative to control (DMSO) (n = 6, ~700 BC cells/z-stack). h Same as in g for proportion of BC cells that invaded above 50 μm. i Left, Velocity of MCF7 in tumor-on-chip in CAF-free (−, gray) or CAF-S4 (blue)-embedded collagen treated or not with DAPT. Each dot is one cell, n ≥ 95/condition. Right, Same as left for median velocity of MCF7 (n = 3). j Same as i for T47D. In all panels, boxplots are median ± 25%–75% quantiles, whisker values range 1.5 × IQR above 75th or below 25th percentiles. Barplots mean ± SEM. a–e p Values from Mann–Whitney test. g–i p Values from paired t-test. At least four CAF-S1 and CAF-S4 tested. Source data provided in Source Data file, with R scripts used.

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