Fig. 5

Modulation of SH3BGRL3 expression impacts on the migration features of MDA-MB-231 cells. (a) Upper: Representative microscopic images of Boyden chamber fields of migrated MDA-MB-231 cells transfected with control scrambled siRNA (CTR) or with SH3BGRL3 specific siRNAs (siRNA #1 or siRNA #2). Lower: Quantification of the migration ability of the same cells as above. Three independent migration experiments were carried out, in duplicate or triplicate. Each chamber slide was divided in nine fields and cell counts/field reported in the graph. For each chamber slide of the scramble controls, the median cell counts/field was set to 100, and data of SH3BGRL3 siRNAs were normalized to their respective scramble control. Field counts for all experiments are gathered in this graph, mean ± SD are reported and unpaired t-test was used for evaluating statistical difference significance. (b) Western blots showing the silencing efficiency of the two SH3BGRL3 specific siRNAs. Below are the SH3BGRL3/GAPDH ratios, normalized to the scramble control ratio, which was set to 100. c) Upper: Representative microscopic images of Boyden chamber fields of migrated MDA-MB-231 cells subjected to lentiviral infection of either control (CTR) or SH3BGRL3 vectors. Lower: Quantification of the migration ability of the same cells as above in two independent migration experiments, each carried out in duplicate. Cell counts and statistical analysis were performed as detailed in a). (d) Western blots showing the efficiency of the over-expression system. Below are the SH3BGRL3/GAPDH ratios, normalized to the control ratio, which was set to 100