Fig. 7: Influence of the 5-LO KO on colony formation, migration and invasion of HCT-116, HT-29 and U-2 OS cells. | Cancer Gene Therapy

Fig. 7: Influence of the 5-LO KO on colony formation, migration and invasion of HCT-116, HT-29 and U-2 OS cells.

From: Knock-out of 5-lipoxygenase in overexpressing tumor cells—consequences on gene expression and cellular function

Fig. 7

A Two dimensional colony formation of the 5-LO KO clones. Cells were seeded into 6-well plates at low density (400 cells/well) and the number of colonies formed after 7 days was assessed by Ponceau red staining. Data are presented as + SD of three independent experiments (Plating efficiency of control vector cells: HCT-116; 72%, HT-29; 70%, U-2 OS; 80%). B Three dimensional colony formation of the 5-LO clones in soft agar. Colonies formed after 3 weeks were stained with crystal violet and counted. U-2 OS cells did not form any colonies under these conditions. Data are presented as mean + SD of three independent experiments (Plating efficiency of control vector cells: HCT-116; 1.6%, HT-29; 0.6%). C Outgrowth/invasion of preformed spheroids into matrigel®. Growth (diameter in µm) was monitored for 14–19 days. Data are presented as mean ± SD of 3–9 independent experiments. D Directed cell migration towards serum was measured in a transwell assay for 3 h (U-2 OS) or 24 h (HCT-116, HT-29). E For investigations on invasive properties of the cells migration through matrigel® was measured. Data on cell migration and invasion in the transwell assay are presented as mean + SD from 3–5 independent experiments. Data depicted in A, B, D, and E are depicted as % vector control. Asterisks indicate significant changes vs. control vector treated cells. *P ≤ 0.05, **P ≤ 0.01, ***P ≤ 0.001.

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