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Fig. 7. Metabolic degradation of Ins(1,4,5)P₃ determines directional responses. In the cartoons, the pipette indicates the stimulated cell type (EC top, VSMC bottom), blue is BAPTA, yellow is 5-PI, orange is XPC with 5-PI and `+' indicates the cell type that is loaded with siRNA. (A,B) BAPTA-loaded VSMCs were stimulated. When 5-PI was loaded into ECs treated with Ins(1,4,5)P₃-R1 siRNA, an increase in EC [Ca²⁺]_i was observed (A) and, when XPC was added to these cells, an increase in EC [Ca²⁺]_i was no longer present (B). (C,D) BAPTA-loaded ECs were stimulated. With the addition of 5-PI to VSMCs, an increase in VSMC [Ca²⁺]_i was observed after EC stimulation (C) and, when XPC was added to the VSMCs with 5-PI, the VSMC Ca²⁺ response to EC stimulation was abolished (D). ^*P<0.05. (E) In transverse sections of the cremasteric VCCC, pores of the Transwell demonstrate the presence of Ins(1,4,5)P₃-R1 (green) and 5-phosphatase (red). The vertical white line indicates the location of the line scan. (F) The line scans correspond to the fluorescent intensity from each of the proteins tested in E. Scale bar: 10 µm.

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