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Epac-1/Rap-1 and its downstream signaling Akt and PLCε, but not Rac-1, mediate cilostazol-induced PGI2 production.

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posted on 2015-07-16, 02:54 authored by Ayako Hashimoto, Michinori Tanaka, Satoshi Takeda, Hideki Ito, Keisuke Nagano

(A) Top, Representative immunoblots showing total Rap-1 expression and Rap-1 activation in HAECs. Middle, Effect of Rap-1-targeting siRNAs (iRap-1) or non-targeting siRNAs (iCt) on protein levels in HAECs. Bottom, Effect of Rap-1 knockdown on PGI2 production in HAECs. HAECs were transfected with iRap-1, or with the iCT. Post-transfection HAECs were treated with vehicle or 30 μM cilostazol (n = 4; † p < 0.05 vs. vehicle t-test). (B) Top, Representative immunoblot showing PLCε expression in HAECs. Middle, Effect of PLCε-targeting siRNAs (iPLCε) or non-targeting siRNAs (iCt) on protein levels in HAECs. Bottom, Effect of PLCε knockdown on PGI2 production in HAECs. HAECs were transfected with iPLCε, or with the iCT. Post-transfection HAECs were treated with vehicle or 30 μM cilostazol (n = 4;† p < 0.05, †† p < 0.01 vs. iCT, t-test). (C) Top, Representative immunoblot showing P-PDK1, PDK1, Akt, and P-Akt expression in HAECs. Bottom, Effect of the PI3Kγ antagonist, AS60520, on PGI2 production in HAECs (n = 4, †† p < 0.01 vs. vehicle, t-test; ** p < 0.01 vs. 30 μM cilostazol alone, lower-tailed Williams’ test).

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