Figure 8.

A and B, relative mRNA expression of IMD normalised against GAPDH (A; similar results were found using β-actin as a housekeeping gene – not shown) or protein level normalised against β-actin (B) are presented as relative to normoxia. n = 3 HMVECs sources run in triplicate: #P < 0.05 vs. normoxia. C–F, viable cell counts for cells which have had the CRLR receptor components knocked-down before exposure to normoxia, ischaemia and ischaemia–reperfusion without/with co-incubation with 10⁻⁹ mol l⁻¹ IMD. n = 3 HAEC sources run in triplicate. *P < 0.05 vs. untreated control, †P < 0.05 vs. non knock-down counterpart, and #P < 0.05 vs. non-IMD counter part. G, blots obtained when membrane protein lysates were immunoprecipitated (IP) with RAMP2 and western blotted with CRLR antibody (and vice versa) to confirm co-precipitation of molecules. The heavy chain of the IP antibody also transfers to the blot and is employed as a loading control. Treatment conditions included no pre-treatment or pre-treatment with either random (non-targeting siRNA) or siRNA directed against CRLR or RAMP2.