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. 2011 Oct 21;52(11):8271–8277. doi: 10.1167/iovs.11-8230

Figure 6.

Figure 6.

Ligand-activated CCR3 coimmunoprecipitates with and activates VEGFR2 and leads to synergistic activation of Rac1. (A) Coimmunoprecipitation of CCR3 and VEGFR2 was performed in CECs incubated with VEGF (20 ng/mL) and CCL1 (10 ng/mL) for 1 hour using antibodies to CCR3 and blotting for VEGFR2 (n = 3). (B) Phospho-VEGFR2 was detected by immunoprecipitation with antibodies against VEGFR2 and a blot for phospho-tyrosine in CECs treated with CCL11 (10 ng/mL) for 0, 15, 30, and 60 minutes; phosphorylation of VEGFR2 in CECs exposed to VEGF (20 ng/mL) for 15 minutes was a positive control. Overall one-way ANOVA P < 0.0001; post-hoc Newman-Keuls multiple comparison testing: *P < 0.05, **P < 0.01, and ***P < 0.001 vs. 0 minutes of CCL11 treatment (n = 3). (C) Rac1 activity in CECs was measured in response to the stimulation of CCL11 (10 ng/mL) and VEGF (20 ng/mL). Rac1 activity in cells with either CCL11 or VEGF treatment alone was used as a control. Overall one-way ANOVA P < 0.0001; post-hoc Newman-Keuls multiple comparison testing: **P < 0.01 and ***P < 0.001 vs. control; #P < 0.05 vs. CCL11 alone; $P < 0.05 vs. VEGF alone (n = 3).