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. Author manuscript; available in PMC: 2020 May 1.
Published in final edited form as: Gynecol Oncol. 2019 Feb 20;153(2):405–415. doi: 10.1016/j.ygyno.2019.01.020

Figure 5. Treatment with G2, a fascin inhibitor, decreases actin bundling, Cdc42-GTP and Rac1-GTP activity in both cancer and stromal cells.

Figure 5.

a. F-actin (Phalloidin-488) and nuclear (Hoechst) staining of cancer cells and primary human mesothelial cells (HPMC) after 24 hour co-culture and subsequent 8 hours of treatment with 50 μM G2 or DMSO (vehicle control). b-e. Cancer cells were cultured on plastic. Twenty-four hours later the cancer cells were treated for 5 hours with 50 μM G2 or DMSO control in serum-free media followed by 10 minute treatment with 50 ng/ml epidermal growth factor (EGF). The levels of GTP-bound Cdc42 or Rac1 were measured using the following assays. b. Cdc42 G-LISA Activation Assay. c. Rac1,2,3 G-LISA Activation Assay. Bars represent mean ± standard error of the mean (n=5 wells per cell type; n=3 independent experiments). Differences between treatments were evaluated using a paired t-test, two-tailed, with p<0.05 (*) and p<0.01 (**).d. Cdc42 Pull-down Activation Assay. e. Immunofluorescent detection of Rac1-GTP (pink). Blue, nuclei. Size bar, 50μm.