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. 2019 Sep 24;11(10):1421. doi: 10.3390/cancers11101421

Figure 6.

Figure 6

3D co-culture tissue reconstructs F331 fibroblasts with either SPARC KD, KD + rSPARC or scr control were suspended into a 3D matrix consisting of methylcellulose + collagen type I and allowed to grow and structure their 3D matrix for 4 days. Cancer cells were seeded onto these reconstructs, incubated for an additional 6 days and IHC stained with a cytokeratin antibody. Typical sections of either F331 (A) or CAFs (B) co-cultured with HCT116 cells used for quantification. Overview of sections can be viewed in corresponding upper panels (size bar = 500 µm) and zoomed images in lower panels (size bar = 100 µm), respectively. For each reconstruct, 5 sections separated by 100µm were analyzed. Results are shown as average invasion depth per section (n = 15) for 3 independent experiments using 2 different siRNAs for both HCT116 and SW480 (C). In addition, reconstructs with CAFs co-cultured with either HCT116 or SW480 (D) were evaluated as well (15 sections, 3 independent experiments for HCT116 and 10 sections, 2 independent experiments, for SW480; 2 different siRNAs for both cell lines). Statistical analysis to evaluate differences between 3 groups were performed by using the Kruskal Wallis test followed by Dunn’s multiple comparisons test, and the Mann Whitney test for 2 groups. *: p < 0.05; **: p < 0.01; Scatter dot plots: center line, means; whiskers, SD; KD, knockdown; rSPARC, human recombinant SPARC; scr, scrambled control.