Fig. 2: Loss of CdGAP results in elevated Rac1-GTP levels in PC-3 cells. | Communications Biology

Fig. 2: Loss of CdGAP results in elevated Rac1-GTP levels in PC-3 cells.

From: CdGAP promotes prostate cancer metastasis by regulating epithelial-to-mesenchymal transition, cell cycle progression, and apoptosis

Fig. 2

a Immunoblot analysis of CdGAP and E-cadherin in human prostate cancer cell lines DU-145, LNCaP, and PC-3. Tubulin was used as a loading control. Graphs provide a densitometry analysis of CdGAP and E-cadherin protein levels represented as the fold change relative to DU-145 cells (n = 3). b mRNA levels of CdGAP (n = 4) and E-cadherin (n = 3) represented as the fold change relative to DU-145 cells. c Immunoblot analysis of CdGAP levels in PC-3 cells infected with scrambled control (shCon) or shRNA targeting CdGAP (shCdGAP). Tubulin was used as a loading control. The graph provides a densitometry analysis of CdGAP protein levels represented as the fold change relative to control (n = 3). d mRNA levels of CdGAP represented as the fold change relative to control (n = 3). e GTP-bound Rac1 was pulled down using GST-CRIB from control (shCon) or CdGAP-depleted PC-3 (shCdGAP) cell lysates. TCL: total cell lysates. Graphs provide a densitometry analysis of GTP-bound Rac1/total Rac1 represented as the fold change relative to control (n = 3). f Control and shCdGAP PC-3 cells were plated on coverslips coated with fibronectin. Actin filaments and nuclei were stained using phalloidin-TRITC and DAPI. Scale bar represents 10 μm. Cell aspect ratio and cell size were quantified (n = 3). shCon: total number of cells = 130; shCdGAP: total number of cells = 166. Two-sample unpaired Student’s t test for comparison between two groups with Welch’s correction in (f). Error bars indicate SEM. ****p < 0.0001 ***p < 0.001; **p < 0.01; *p < 0.05.

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