Chunk #60 — Experimental Procedures — HEK cell transfection

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Posttranscriptional regulation of BK channel splice variant stability by miR-9 underlies neuroadaptation to alcohol.

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HEK293 were cultured in DMEM medium with 10% fetal bovine serum and, for transfection, confluent HEK293 cells were plated on 60 mm Petri dishes and transfected using PolyFect transfection reagent (Qiagen,Balencia, CA) complexed with BK variants (BK-Insertless, BK-ALCOREX, BK-STREX) in pVAX vector (Invitrogen) cDNAs.