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. 2015 Jan 7;7(1):96–111. doi: 10.3390/cancers7010096

Figure 4.

Figure 4

Figure 4

The Role of Akt3 in Src-Induced Podosome and Rosette Formation. (A) Western blots showing the efficiency of shRNA knockdown of Akt3 compared to negative shRNA control. Src (Y527F) cells were transduced with 3 different shRNAs targeting different regions of Akt3 (Akt3-shRNA1, Akt3-shRNA2 or Akt3-shRNA3). GAPDH was used as a loading control; (B) Cells containing podosomes and/or rosettes, rosettes, and those with >50 podosomes per cell were counted. Error bars represent standard deviation from 3 separate experiments and * represents p-value < 0.05 with respect to control cells; (C) Cells expressing Akt3-shRNA3 were transiently transfected with Akt1 siRNA and or Control siRNA. Cells were counted to determine the relative number of cells displaying podosomes and/or rosettes, rosettes, and those with >50 podosomes per cell. Error bars represent standard deviation from three separate experiments and * represents p-value < 0.05 with respect to control siRNA; (D) Cells were seeded on fibronectin substrate for 20 h, and areas of digestion were measured. Error bars represent standard deviation from three separate experiments and * represent p-value < 0.05; (E) Representative images of cells on fibronectin substrates are shown. F-actin was stained green with FITC-phalloidin and fibronectin immune-stained with TRITC-antibody (red). Scale bars represents 20 μm.