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

Figure 4. HSP70 overexpression and co-distribution with VEGFR2.

Figure 4

(A) Lentiviral transduced HUVECs expressing human HSP70-FLAG were labeled for VEGFR2 (green) and HSP70-FLAG (red), which were detected using labeled secondary antibodies and co-distribution shown (yellow). Bar, 10 µm. (B) Quantification of VEGFR2 co-distribution with HSP70-FLAG under control or HSP70 overexpression conditions. Error bars denote ±SEM (n = 15), ***p<0.005 using Student’s t-test. (C) HEK-293T cells were transfected with VEGFR2 and/or HSP70-FLAG and processed for immunoblotting (IB) using anti-VEGFR2 antibody. Arrowhead denotes mature transfected VEGFR2. (D) HUVECs were treated ±geldanamycin (4 h) and immunoprecipitation (IP) carried out for control protein (Goat IgG) and VEGFR2 as per Figure 2. Isolated protein complexes were processed for immunoblotting (IB) using anti-VEGFR2 and anti-CHIP antibodies. Arrrowhead denotes mature VEGFR2. (E) HUVECs were treated with either control siRNA or siRNA duplex targeted against CHIP and treated ±geldanamycin (4 h). Cells were lysed and processed for imunoblotting (IB). Representative immunoblots shown. CHIP, carboxy terminus of HSP70-interacting protein.