Figure 2.
Loss of ARHGAP17 expression promotes invasion. (A) Representative micrographs of SUM159 cells degrading a fluorescent-gelatin/collagen IV substrate. F-actin staining was used as a marker for invadopodia. The zoomed panel on the right shows a cluster of invadopodia colocalizing with degradation spots (scale bar for left and right panels, 10 μm). (B) Representative micrographs of F-actin–stained SUM159 cells degrading fluorescent-gelatin/collagen IV matrix after 24 h PDBu treatment (scale bar, 20 μm). (C) Quantification of matrix degradation by SUM159 cells. Results are expressed as mean ± SEM of three independent experiments in which at least 20 random FOVs per condition per experiment were measured. Significance was tested with a two-tailed t test: **, P < 0.01. (D) Micrographs of SUM159 spheroids embedded in Collagen I matrix and treated with mitomycin C to inhibit cell proliferation and with either DMSO or GM6001 to inhibit metalloprotease activity (scale bar for both panels, 50 μm). (E) Quantification of spheroid invasion 48 h after embedding in Collagen I matrix. Results are expressed as mean ± SD of at least six spheroids per condition. Significance was tested with a two-tailed t test: *, P < 0.05. (F) Micrographs of CTRL, ARHGAP17 KO, and Rescue SUM159 spheroids embedded in Collagen I matrix to promote invasion and treated with mitomycin C to inhibit cell proliferation (scale bar for both panels, 50 μm). (G) Quantification of spheroid invasion 48 h after embedding in Collagen I matrix. Results are expressed as mean ± SD of three independent experiments in which at least four spheroids per condition per experiment were measured. Significance was tested with a two-tailed t test: *, P < 0.05.

Loss of ARHGAP17 expression promotes invasion. (A) Representative micrographs of SUM159 cells degrading a fluorescent-gelatin/collagen IV substrate. F-actin staining was used as a marker for invadopodia. The zoomed panel on the right shows a cluster of invadopodia colocalizing with degradation spots (scale bar for left and right panels, 10 μm). (B) Representative micrographs of F-actin–stained SUM159 cells degrading fluorescent-gelatin/collagen IV matrix after 24 h PDBu treatment (scale bar, 20 μm). (C) Quantification of matrix degradation by SUM159 cells. Results are expressed as mean ± SEM of three independent experiments in which at least 20 random FOVs per condition per experiment were measured. Significance was tested with a two-tailed t test: **, P < 0.01. (D) Micrographs of SUM159 spheroids embedded in Collagen I matrix and treated with mitomycin C to inhibit cell proliferation and with either DMSO or GM6001 to inhibit metalloprotease activity (scale bar for both panels, 50 μm). (E) Quantification of spheroid invasion 48 h after embedding in Collagen I matrix. Results are expressed as mean ± SD of at least six spheroids per condition. Significance was tested with a two-tailed t test: *, P < 0.05. (F) Micrographs of CTRL, ARHGAP17 KO, and Rescue SUM159 spheroids embedded in Collagen I matrix to promote invasion and treated with mitomycin C to inhibit cell proliferation (scale bar for both panels, 50 μm). (G) Quantification of spheroid invasion 48 h after embedding in Collagen I matrix. Results are expressed as mean ± SD of three independent experiments in which at least four spheroids per condition per experiment were measured. Significance was tested with a two-tailed t test: *, P < 0.05.

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