Fig. 1.

Nitro-oleate increased proliferation and migration of LLc1 cells through inhibition of sEH and EET signalling

(A) 5 μM or 10 μM nitro-oleate reduced sEH activity. In contrast, sEH activity was not reduced by the non-electrophile oleate (N=12 per group from 3 independent experiments). The time graph shows that sEH-dependent hydrolysis of PHOME to 6-methoxy-2-naphtaldehyde (6M2N), which is fluorescent, is linear over 60 min. (B-D) Real-time monitoring of LLc1 cell proliferation (cell index) and proliferation rates calculated from these measurements. (B) 5 μM or 10 μM nitro-oleate stimulated proliferation compared to untreated or DMSO-treated control cells (N=4 independent experiments). (C) In addition, oleate did not stimulate cell proliferation (N=4 independent experiments). (D) The increased proliferation induced by 5 μM nitro-oleate was normalized by the EET antagonist 14,15-EE-5(Z)-E (N=4 independent experiments). (E) Real-time monitoring of LLc1 cells trans-well migration (cell index) and migration rates calculated from these measurements. 5 μM nitro-oleate stimulated transmigration compared to untreated or vehicle (DMSO)-treated control cells (N=4 independent experiments). *P < 0.05 significantly different compared to untreated control. #P < 0.05 significantly different compared to the nitro-oleate treated group.