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. Author manuscript; available in PMC: 2019 Jan 15.
Published in final edited form as: Clin Cancer Res. 2017 Dec 5;24(2):420–432. doi: 10.1158/1078-0432.CCR-17-1776

Figure 5. Vimentin regulates CAF invasion and stroma-cancer cell crosstalk.

Figure 5

(A) Western blot demonstrating generation of shVIM CAFs with two different shRNA clones. (B) Immunofluorescence of vimentin in pLKO.1 andshVIM19 CAFs (scale=50μm). (C) Immunofluorescence of actin and vimentin in pLKO.1 and shVIM CAFs demonstrating cell size (10× scale= 100μm, 20× scale = 100μm). (D) Quantification of cell area of pLKO.1 and shVIM CAFs. Significance determined by unpaired t-test (p<0.05). (E) Representative images of spheroid invasion assay with pLKO.1 and shVIM CAFs in matrigel (scale = 100μm). (F) Quantification of invasive area in spheroid invasion assay. Significance determined by unpaired t-test (p<0.05). (G) Representative images ofcoculture spheroid invasion assay with H460 lung cancer line alone or in combination with pLKO.1 or shVIMCAFs (scale = 100μm). (H) Circularity of invasive areas in representative trial (3 total trials) of coculture spheroids with pLKO.1(n=6) or shVIM (n=3) CAFs. Value of 1= perfect circle. Significance determined by unpaired t-test (p<0.05). (I) Quantification of invasive chains in representative coculture spheroids with pLKO.1 or shVIM CAFs. Significance determined by unpaired t-test (p<0.05). (J) Representative images of immunofluorescence of FSP1and phalloidin in coculture spheroids. Yellow arrows mark CAFs positive for cytoplasmic FSP1 leading invasive chain of H460s (scale = 100μm).