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. 2011 Nov 18;302(3):H724–H732. doi: 10.1152/ajpheart.00739.2011

Fig. 5.

Fig. 5.

p66Shc is involved in VEGF-induced VEGFR2 autophosphorylation in caveolae/lipid rafts in ECs. A: sucrose gradient centrifugation was performed to isolate caveolae/lipid rafts (C/LR), equal volume of each fraction from top to bottom was IB with anti-VEGFR2, Nox2, caveolin-1, and paxillin (marker for non-C/LR) Abs. C/LR fraction were appeared in fractions 4–6. B, left: HUVECs were stimulated with or without VEGF (20 ng/ml) for 5 min, and followed by C/LR fractionation. Equal amounts of proteins from C/LR fraction were IB with anti-VEGFR2-pY1175, total VEGFR2, and caveolin-1 Abs (n = 3). B, right: HUVECs transfected with control or p66Shc siRNAs were stimulated with VEGF (20 ng/ml) for 5 min, and the C/LR fraction were IB with anti-VEGFR2-pY1175, total VEGFR2, and caveolin-1 Abs. B, bottom: averaged data, expressed as VEGF-induced fold change over basal in control siRNA-treated groups (means ± SE). *P < 0.05.