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. 2006 Mar 15;395(Pt 1):65–71. doi: 10.1042/BJ20050582

Figure 6. EGF-induced CD44 cleavage and CD44-dependent cell migration are mediated by ADAM10.

Figure 6

(A) Effect of ADAM10 siRNA on EGF-induced CD44 cleavage. U251MG cells were transfected with ADAM10 siRNA (lane 3) or control siRNA (lane 4), and left untreated (lane 1) or treated with 10 ng/ml EGF for 2 h (lanes 2–4). CD44 cleavage product was detected by Western blot analysis (top panel). Blocking of endogenous ADAM10 expression by siRNA was monitored by Western blot analysis using anti-ADAM10 pAb (middle panel). β-Tubulin was detected as an internal control (bottom panel). The histogram shows the results of quantitative analysis of the CD44 cleavage product bands as means±S.D. for three independent experiments. (B) Effects of EGF on the cell migration on hyaluronan. The U251MG cells transfected with ADAM10 siRNA (bars 5 and 6) or control siRNA (bars 7 and 8) or the cells left untransfected (bars 1–4) were pre-incubated in the presence (bars 2, 4, 6 and 8) or absence (bars 1, 3, 5 and 7) of 10 μg/ml BRIC235 mAb for 10 min, and then incubated at 37 °C for 24 h in the absence (bars 1 and 2) or presence (bars 3–8) of 10 ng/ml EGF in the hyaluronan-coated Transwell chambers. Results are mean ratios of the control migration, expressed as means±S.D. for triplicate determinations. (C) Effects of EGF on the cell migration on fibronectin. The U251MG cells transfected with ADAM10 siRNA (bar 3) or control siRNA (bar 4) or the cells left untransfected (bars 1–2) were incubated for 24 h in the absence (bar 1) or presence (bars 2–4) of 10 ng/ml EGF in the fibronectin-coated Transwell chambers. Results are mean ratios to the control migration, expressed as means±S.D. for triplicate determinations. *, P<0.01 compared with untreated cells; †, P<0.01 compared with only EGF-treated cells (ANOVA with Dunnett's post hoc test).