Sigenome non targeting sirna 5

ON-TARGETplus SMARTpool siRSK1, siRSK2, and siGENOME Non-Targeting siRNA #5 (siRNA negative control) were purchased from Thermo-Scientific.

Using a modification of methods previously described [14] (link), PAECs were transfected with non-targeting (control) oligonucleotides (siGENOME Non-targeting siRNA #5) or SNAT1 targeting oligonucleotides (sense, 5′-ACGAACAGCCAUUUGGAAAUU-3′; anti-sense, 5′-UUUCCAAAUGGCUGUUCGUUU-3′; sense, 5′-CCGGAAGAUGAUAACAUUAUU-3′ anti-sense, 5′-UAAUGUUAUCAUCUUCCGGUU-3′; anti-sense, 5′-UAAUGUUAUCAUCUUCCGGUU-3′; sense, 5′-CAGUAACACUUCUGUCUAUUU-3′ anti-sense, 5′-AUAGACAGAAGUGUUACUGUU-3′) purchased from Thermo Scientific. For transfection, EGM-2 was replaced with OPTI-MEM (Gibco). PAECs were transfected with a 300–600 pmol suspension of either non-targeting or SNAT1 oligonucleotides (100–200 pmol for each of the 3 different SNAT1 targeting oligonucleotides) using lipofectamine (Invitrogen) in OPTI-MEM. After 4 hours, the medium was replaced with EGM-2 and 10% FBS.