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. 2018 Sep 17;12(10):1735–1752. doi: 10.1002/1878-0261.12367

Figure 4.

Figure 4

ABCA1 overexpression favours spheroid formations and promotes invasion in a three‐dimensional model. (A) Representative images of cell growth in a spheroid formation assay in basement membrane matrices (Matrigel™). Scale bar corresponds to 50 μm. Growth of DLD1 spheres is represented in the middle panel. The graph presents the average of 30 different spheres. The percentages of cells that were positive for histone H‐3 phosphorylation are presented in the dot plot graph. n = 3; 10 pictures per condition were taken and quantified. The values in the histograms correspond to mean ± SEM. The significance of differences between experimental groups was determined by t‐test. DLD1_NoORF: 3.311 ± 0.6882; DLD1_ABCA1: 5.736 ± 0.7247. (B) The percentage of spheroids that present protrusions is shown in the left‐hand panel. The values in the histograms correspond to mean ± SEM. ANOVA analysis was performed to determine the significance of differences between groups (F (4,9) = 6.146; P = 0.0115). In the dot plot graph, quantification of the longest protrusions per spheroid are represented. A t‐test analysis was used to determine the significance of the analysis. DLD1_NoORF: 3.914 ± 3.037; DLD1_ABCA1: 20.92 ± 6.231. The experiment was performed three times; 10–20 spheroids were quantified. Scale bars correspond to 100 μm. (C) Levels of expression of E‐cad and Vim mRNA. The values in the histograms correspond to mean ± SEM. The significance of differences between experimental groups was determined by t‐test. For E‐cad, DLD1_NoORF: 1.003 ± 0.003161; DLD1_ABCA1: 0.6925 ± 0.02835. For Vim, DLD1_NoORF: 0.9942 ± 0.009214; DLD1_ABCA1: 1.515 ± 0.07528.