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. 2012 Nov 6;7(11):e48539. doi: 10.1371/journal.pone.0048539

Figure 3. HSP70 requirement for geldanamycin-stimulated VEGFR2 degradation.

Figure 3

(A) HUVECs pre-treated with siRNA duplex ‘a’ to HSP70 alone or with geldanamycin (4 h) were analyzed by immunoblotting (IB) for proteins as indicated. Arrowhead indicates mature VEGFR2. (B) Quantification of mature VEGFR2 levels under conditions in panel A. (C) Quantification of HSP70 levels in HUVECs pre-treated with either of two different synthetic siRNA duplexes ‘a’ or ‘b’ to HSP70±geldanamycin. (D) Quantification of VEGFR2 levels in HSP70-depleted HUVECs (duplex ‘a’ or ‘b’) ±geldanamycin (4 h). Error bars denote ±SEM (n≥3), *p<0.05; **p<0.01; ***p<0.005 using one-way ANOVA. (E) HUVECs pre-treated with siRNA duplexes for HSP70 or control siRNA were treated ±geldanamycin (4 h) ±VEGF-A (5 min) and analyzed for signaling events by immunoblotting. (F) Quantification of phospho-PLCγ1 levels from panel E. Error bars denote ±SEM (n≥3), *p<0.05 using one-way ANOVA.