Fig. 5

Decreased Rho guanine nucleotide dissociation inhibitor (RhoGDI2) levels correlate with increased cell motility and poor outcome in human epidermal growth factor receptor 2 (HER2) + breast cancers. a RhoGDI2/ARHGDIB mRNA expression levels were assessed in HER2+ breast tumors curated in publicly available breast cancer expression array datasets, using Km Plotter software (kmplot.org) to identify Affymetrix probe 1555812_a_at (ARHGDIB/RhoGDI2; N = 150). Samples were separated at the median level RhoGDI2/ARHGDIB expression, and plotted against metastasis-free survival for each sample in the Kaplan-Meier curve shown. b-d Cells were transiently transfected with siRNA sequences against RhoGDI2 or non-specific control siRNA. Whole cell lysates or Pak-Pak binding domain pull-downs from whole cell lysates were assessed by western analysis (b). Cells were assessed for invasion and migration through Matrigel-coated transwells 24 h after seeding by crystal violet staining. Representative images are shown (c). Quantitation of cells on the lower side of the filter is shown (d). Midlines are the average of three repeats and points are replicates, with each assessed in duplicate. Student’s t test was used to assess significance. HR hazard ratio, shScr scrambled short hairpin shRNA sequences, GTP guanosine triphosphate, shRictor short hairpin RNA sequences against Rictor, siCtrl small interfering control