Figure 1.
ARHGAP17 negatively regulates invadopodia. (A) Representative micrographs of a SUM159 cell with an invadopodia cluster. Cells were treated with PDBu for 30 min and stained for cortactin and F-actin (scale bar for both panels, 5 μm). (B) shRNA screen of candidate RhoGAPs in SUM159 cells. Cells stably expressing the indicated shRNAs were treated with PDBu for 30 min and stained for cortactin and F-actin as invadopodia markers. Colors represent different RhoGAPs, and individual bars represent unique shRNA targeting sequences. Results are expressed as the percentage of cells with invadopodia from four FOVs per condition. (C) Western blot of ARHGAP17 expression in SUM159 cells stably expressing non-targeting and ARHGAP17-specific shRNAs. Tubulin was used as a loading control. (D) Percentage of cells with invadopodia in CTRL or ARHGAP17 KD SUM159 cells treated with PDBu for 30 min. Results are expressed as mean ± SEM of three independent experiments in which at least 200 cells per condition per experiment were counted. Significance was tested with a two-tailed t test: ***, P < 0.001. (E) Western blot of CTRL, ARHGAP17 KD, and Rescue with shRNA-resistant myc-ARHGAP17 in SUM159 cells. (F) Percentage of cells with invadopodia in CTRL, ARHGAP17 KD, and Rescue SUM159 cells treated PDBu for 30 min. Results are expressed as mean ± SEM of three independent experiments in which at least 200 cells per condition per experiment were counted. Significance was tested with a two-tailed t test: **, P < 0.01. (G) Representative micrographs of CTRL, ARHGAP17 KD, and Rescue with myc-ARHGAP17 in SUM159 cells. F-actin and cortactin staining were used as markers for invadopodia while myc staining was used to visualize myc-ARHGAP17. Cells were treated with PDBu for 30 min before fixation and staining. Arrowheads highlight invadopodia clusters and the zoomed region is indicated with a dashed line (scale bar, 20 μm; zoom scale bar, 5 μm). For all blots, molecular weights are indicated in kD. Source data are available for this figure: SourceData F1.

ARHGAP17 negatively regulates invadopodia. (A) Representative micrographs of a SUM159 cell with an invadopodia cluster. Cells were treated with PDBu for 30 min and stained for cortactin and F-actin (scale bar for both panels, 5 μm). (B) shRNA screen of candidate RhoGAPs in SUM159 cells. Cells stably expressing the indicated shRNAs were treated with PDBu for 30 min and stained for cortactin and F-actin as invadopodia markers. Colors represent different RhoGAPs, and individual bars represent unique shRNA targeting sequences. Results are expressed as the percentage of cells with invadopodia from four FOVs per condition. (C) Western blot of ARHGAP17 expression in SUM159 cells stably expressing non-targeting and ARHGAP17-specific shRNAs. Tubulin was used as a loading control. (D) Percentage of cells with invadopodia in CTRL or ARHGAP17 KD SUM159 cells treated with PDBu for 30 min. Results are expressed as mean ± SEM of three independent experiments in which at least 200 cells per condition per experiment were counted. Significance was tested with a two-tailed t test: ***, P < 0.001. (E) Western blot of CTRL, ARHGAP17 KD, and Rescue with shRNA-resistant myc-ARHGAP17 in SUM159 cells. (F) Percentage of cells with invadopodia in CTRL, ARHGAP17 KD, and Rescue SUM159 cells treated PDBu for 30 min. Results are expressed as mean ± SEM of three independent experiments in which at least 200 cells per condition per experiment were counted. Significance was tested with a two-tailed t test: **, P < 0.01. (G) Representative micrographs of CTRL, ARHGAP17 KD, and Rescue with myc-ARHGAP17 in SUM159 cells. F-actin and cortactin staining were used as markers for invadopodia while myc staining was used to visualize myc-ARHGAP17. Cells were treated with PDBu for 30 min before fixation and staining. Arrowheads highlight invadopodia clusters and the zoomed region is indicated with a dashed line (scale bar, 20 μm; zoom scale bar, 5 μm). For all blots, molecular weights are indicated in kD. Source data are available for this figure: SourceData F1.

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