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. Author manuscript; available in PMC: 2020 May 1.

Published in final edited form as: Gynecol Oncol. 2019 Feb 20;153(2):405–415. doi: 10.1016/j.ygyno.2019.01.020

Figure 4. — a. Boyden chamber migration assay for HeyA8, Ovcar5, and Tyk-nu ovarian cancer cells (left panel), primary human cancer-associated fibroblasts (CAF; middle panel) and primary human omental mesothelial cells (HPMC; right panel) treated with 50 μM G2 or DMSO control. (n = 5 fields per well, n=3 wells per cell type; n=3 independent experiments). b. Boyden chamber migration assay for HeyA8 ovarian cancer cells transfected with control or fascin siRNA and treated with 50 μM G2 or DMSO control. c. Wound healing assay for HeyA8, Ovcar5, and Tyk-nu ovarian cancer cells (left panel), primary human cancer-associated fibroblasts (CAF; middle panel) and primary human omental mesothelial cells (HPMC; right panel) treated with 50 μM G2 or DMSO control. (n=5 wells per cell type; n=3 independent experiments). d. Oris plug migration assay for HeyA8 cells co-cultured with cancer-associated fibroblasts (CAF) and treated with 10 μM G2 or DMSO control for 72 hours. (n=6 wells per treatment group; data shown is representative of 2 independent experiments). Representative fluorescent images of GFP-labeled HeyA8 ovarian cancer cell migration at 0, 24, and 72 hours. e. Wound healing assay for HeyA8, Ovcar5, and Tyk-nu ovarian cancer cells in co-culture with primary human omental mesothelial cells (HPMC) treated with 50 μM G2 or DMSO control. (n=5 wells per cell type; n=3 independent experiments). Representative fluorescent images 24h after wound healing assay of GFP-labeled ovarian cancer cells and unlabeled primary human mesothelial cells. f. Ex-vivo primary human omental tissue colonization assay using HeyA8, Ovcar5, and Tyk-nu ovarian cancer cells for 16 hours with 50μM G2 or DMSO control. (n=5 tissues from same patient per cell type; n=2 independent experiments). Bars represent mean ± standard error of the mean. Differences between treatments were evaluated using a paired t-test, two-tailed, with p<0.01 (**), p<0.001 (***), p<0.0001 (****).

Figure 4. — a. Boyden chamber migration assay for HeyA8, Ovcar5, and Tyk-nu ovarian cancer cells (left panel), primary human cancer-associated fibroblasts (CAF; middle panel) and primary human omental mesothelial cells (HPMC; right panel) treated with 50 μM G2 or DMSO control. (n = 5 fields per well, n=3 wells per cell type; n=3 independent experiments). b. Boyden chamber migration assay for HeyA8 ovarian cancer cells transfected with control or fascin siRNA and treated with 50 μM G2 or DMSO control. c. Wound healing assay for HeyA8, Ovcar5, and Tyk-nu ovarian cancer cells (left panel), primary human cancer-associated fibroblasts (CAF; middle panel) and primary human omental mesothelial cells (HPMC; right panel) treated with 50 μM G2 or DMSO control. (n=5 wells per cell type; n=3 independent experiments). d. Oris plug migration assay for HeyA8 cells co-cultured with cancer-associated fibroblasts (CAF) and treated with 10 μM G2 or DMSO control for 72 hours. (n=6 wells per treatment group; data shown is representative of 2 independent experiments). Representative fluorescent images of GFP-labeled HeyA8 ovarian cancer cell migration at 0, 24, and 72 hours. e. Wound healing assay for HeyA8, Ovcar5, and Tyk-nu ovarian cancer cells in co-culture with primary human omental mesothelial cells (HPMC) treated with 50 μM G2 or DMSO control. (n=5 wells per cell type; n=3 independent experiments). Representative fluorescent images 24h after wound healing assay of GFP-labeled ovarian cancer cells and unlabeled primary human mesothelial cells. f. Ex-vivo primary human omental tissue colonization assay using HeyA8, Ovcar5, and Tyk-nu ovarian cancer cells for 16 hours with 50μM G2 or DMSO control. (n=5 tissues from same patient per cell type; n=2 independent experiments). Bars represent mean ± standard error of the mean. Differences between treatments were evaluated using a paired t-test, two-tailed, with p<0.01 (**), p<0.001 (***), p<0.0001 (****).