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. 2019 May 30;11(6):758. doi: 10.3390/cancers11060758

Figure 4.

Figure 4

SEMA7A (Semaphorin7A) and Glioma Associated Stem Cells (GASC)-derived exosomes activate FAK signalling on glioma stem cells. (A,B) Glioma Stem Cells (GSC) were treated with either recombinant SEMA7A-Fc for 2, 5 and 10 min at 10 ng/mL or 100 ng/mL and paired GASC-derived exosomes (10 µg/mL) for 5, 10 and 30 min (C,D). Total cells lysates were resolved on 10% Sodium Dodecyl Sulphate PolyAcrylamide Gel Electrophoresis (SDS-PAGE) and blotted to identify the phosphorylated and non-phosphorylated form of focal adhesion kinase protein (FAK) (A,C). (B,D) Densitometric analysis was performed using the software available in the Gel-doc instrument (Alliance Uvitec, Ltd. Cambridge, UK) to quantify the level of FAK (Focal Adhesion Kinase) phosphorylation. Histograms represent results reported as fold change of p-FAK of treated vs. untreated cells (Ctrl). Values were calculated as the ratio of IOD (Integrated Optical Density) pFAK/FAK after normalisation on beta actin level. Data are presented as mean ± standard deviation (n = 4). *** p < 0.001 vs. Ctrl; * p < 0.05 vs. Ctrl. Statistical analysis was performed by repeated measurements one-way Anova followed by Bonferroni Multiple Comparison post-test (GraphPad Prism 5).