Figure 3.
ARHGAP17 knockdown increases invadopodia stability and accumulation. (A) Representative micrographs of CTRL and ARHGAP17 KO SUM159 cells stained for F-actin (scale bar, 10 μm) and quantification of percent invadopodia coverage after 30 min PDBu treatment. White dashed lines represent the cell outline and yellow dashed lines represent invadopodia clusters. Results are expressed as mean ± SEM of three independent experiments in which at least 30 cells per condition per experiment were counted. Significance was tested with a two-tailed t test: *, P < 0.05. (B) Time course of percentage of SUM159 cells (CTRL, ARHGAP17 KD, and Rescue) forming invadopodia after PDBu treatment at different intervals. Results are expressed as mean ± SEM of three independent experiments in which at least 200 cells per condition per experiment were counted. (C) Example of single invadopodia dynamics showing assembly, stability, and disassembly phases as indicated by measuring TKS5-mTagBFP2 intensity over time at 30 s intervals after PDBu treatment. The gray line represents a second-order smoothing polynomial. (D) Representative micrographs of single invadopodia dynamics of SUM159 cells expressing TKS5-mTagBFP2 as a marker for invadopodia after PDBu treatment (scale bar, 1 μm). (E–G) Quantification of invadopodia dynamics. (E) Invadopodia lifetime (min). (F) Invadopodia stability (min). (G) Comparison of average assembly, stability, and disassembly time in CTRL, ARHGAP17 KO, and Rescue SUM159 cells. In E and F, results are expressed as mean ± SEM of three independent experiments in which at least 12 invadopodia per condition per experiment were measured. In E and F, large circles represent biological replicates, small circles represent technical replicates. Significance was tested with a two-tailed t test: *, P < 0.05; **, P < 0.01; ***, P < 0.001.

ARHGAP17 knockdown increases invadopodia stability and accumulation. (A) Representative micrographs of CTRL and ARHGAP17 KO SUM159 cells stained for F-actin (scale bar, 10 μm) and quantification of percent invadopodia coverage after 30 min PDBu treatment. White dashed lines represent the cell outline and yellow dashed lines represent invadopodia clusters. Results are expressed as mean ± SEM of three independent experiments in which at least 30 cells per condition per experiment were counted. Significance was tested with a two-tailed t test: *, P < 0.05. (B) Time course of percentage of SUM159 cells (CTRL, ARHGAP17 KD, and Rescue) forming invadopodia after PDBu treatment at different intervals. Results are expressed as mean ± SEM of three independent experiments in which at least 200 cells per condition per experiment were counted. (C) Example of single invadopodia dynamics showing assembly, stability, and disassembly phases as indicated by measuring TKS5-mTagBFP2 intensity over time at 30 s intervals after PDBu treatment. The gray line represents a second-order smoothing polynomial. (D) Representative micrographs of single invadopodia dynamics of SUM159 cells expressing TKS5-mTagBFP2 as a marker for invadopodia after PDBu treatment (scale bar, 1 μm). (E–G) Quantification of invadopodia dynamics. (E) Invadopodia lifetime (min). (F) Invadopodia stability (min). (G) Comparison of average assembly, stability, and disassembly time in CTRL, ARHGAP17 KO, and Rescue SUM159 cells. In E and F, results are expressed as mean ± SEM of three independent experiments in which at least 12 invadopodia per condition per experiment were measured. In E and F, large circles represent biological replicates, small circles represent technical replicates. Significance was tested with a two-tailed t test: *, P < 0.05; **, P < 0.01; ***, P < 0.001.

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