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[Preprint]. 2023 Mar 20:2023.02.24.529926. Originally published 2023 Feb 24. [Version 2] doi: 10.1101/2023.02.24.529926

Figure 3. Fibroblast phenotype and activation in lung hydrogels in breast cancer conditioned media.

Figure 3.

a) Representative fluorescent images of HLFs cultured in 3D lung hydrogels with breast cancer conditioned media showing α-SMA (orange), and FAP (green) expressions along with merged HLF images with nuclei staining with DAPI (blue). Scale bar: 100 μm. b) Quantification of α-SMA expression from HLFs cultured in lung gels with breast cancer conditioned media MDA-MB-231 CM, MDA-MB-231 LM2 CM and BT474 CM along with controls. c) Quantification of FAP expression from HLFs cultured in lung gels with the different media conditions. d) Correlation between α-SMA and FAP expressions from HLFs cultured in lung gels with the different media. e) Fold change in cell count for HLFs cultured in lung gels with the different media conditions. f) Percentage of proliferative Ki67+ cells in HLFs cultured in lung gels with the different media conditions. g) Cell area for HLFs cultured in lung gels the different media conditions. h) Cell circularity for HLFs cultured in lung gels with the different media conditions. i) Quantification of number of non-activated and activated cells migrated towards breast cancer conditioned media in trans-well cell migration assay. j) Quantification of number of breast cancer cells migrated towards non-activated and activated HLF conditioned media. All data are mean + s.d. Statistical analyses were performed using Prism (GraphPad). Data in (b), (c), (e), (f), (g), (h), (i) and (j) were analyzed using a one-way analysis of variance followed by a Dunnett’s multiple comparison test with 95% confidence interval. *, **, ***, and **** indicate P < 0.05, P < 0.01, P < 0.001, and P < 0.0001.