Fig 1: Changes in the amounts of total PA in COS-7 cells by overexpressing SMSr.A, human SMSr-V5 was overexpressed in COS-7 cells. After 24 h of transfection, the cells were lysed and immunoprecipitated with an anti-V5 antibody (MA5-15253, Invitrogen), followed by immunoblotting to confirm the expression of SMSr-V5 using the anti-V5 antibody. B, total PA levels in COS-7 cells transfected with vector alone or SMSr-V5 were measured using LC-MS/MS (30). The values are presented as the mean ± S.D. (n = 6). *p < 0.05.
Fig 2: PAP activity of SMSr expressed in mammalian cells.A, SMSr-V5 and SMSr-D348E were overexpressed in COS-7 cells. After 24 h of transfection, the cells were lysed and immunoprecipitated with an anti-V5 antibody (MA5-15253, Invitrogen), followed by immunoblotting to confirm the expression of SMSr-V5 using the anti-V5 antibody. B, PAP, C, PI-PLC, and D, CPES activities in immunoprecipitates were measured by quantitation of 16:0/18:1-DG or d18:1/18:0-CPE levels using LC-MS/MS. The values are presented as the mean ± S.D. (n = 3). ∗p < 0.05 (versus SMSr-V5).
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