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. 2016 Apr 8;7(32):50972–50985. doi: 10.18632/oncotarget.8646

Figure 6. E7 down-regulation by siRNA hinders the acquisition of the metastatic phenotype.

Figure 6

(A) Left panel. Western blot analysis of vimentin (VIM), E-cadherin (E-CAD) on C-33A, SiHa and Caski cells transfected with control siRNA and E7 siRNA. Loading control was evaluated using an anti-GAPDH MAb. (B) Migration test performed by scratch assay on C-33A, SiHa and Caski cells transfected with control siRNA or with E7siRNA. Images were captured by phase-contrast microscopy using a 4.6 × objective at 0, 24/72 h post wounding. A representative experiment among three is shown. (C) Left panel Quantification of the area reduction represented as the percentage of wounded area at time 0. (*) Indicates p < 0.01 between cells transfected with control siRNA and E7 siRNA in the indicated sample. Central panel. Invasion test on C-33A, SiHa and CaSki cell lines performed in vitro by using transwell culture inserts (8.0-μm pore size) coated with Matrigel after E7 silencing by siRNA. Data are reported as mean ± SD of the percentage of invading cells obtained in three independent experiments. Right panel. Flow cytometry evaluation of F-actin intracellular content (expressed by the median fluorescence intensity) in control siRNA-transfected or in E7 siRNA-transfected cells. Results are reported as mean ± SD obtained from three independent experiments. (*) Indicates P < 0.01 between cells transfected with control siRNA and E7 siRNA in the indicated samples.