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. 2015 Sep 30;6(31):31413–31427. doi: 10.18632/oncotarget.5105

Figure 5. CLIC1 secreted protein resides in EVs released from GBM CSCs.

Figure 5

A. Representative immunoblots showing the expression of CLIC1 in three different GBM CSCs and in their respective media. GAPDH was used as loading control. B. Size profile of GBM CSC-derived EVs measured by NTA. Shown is the profile from a representative experiment. C. hGBM#10 CSC whole cell lysates (WCL) and lysates from corresponding EVs were analysed for the indicated proteins by Western blotting. A representative immunoblot is shown. D. Whole cell lysates (WCL) obtained from control hGBM#10 CSCs (NT) and CLIC1 silenced hGBM#10 CSCs (siCLIC1), as well as lysates of corresponding EVs, were analysed by Western blotting using antibodies against CLIC1, and the EV markers CD63 and tsg101. A representative immunoblot is shown. E. hGBM#10 CSCs were cultured in presence of EVs (50 μg/ml) derived from either NT or siCLIC1 hGBM#10 CSCs for 120 hours and assessed for cell growth by 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay. Three independent experiments were performed; error bars represent standard error; *p < 0.05.