THP-1 cells were cultured in RPMI-1640 (Biosera) supplemented with 10% fetal calf serum (FCS) (Biosera) and 1% penicillin-streptomycin in a humidified incubator at 37°C containing 5% CO2. 5 × 105 THP-1 cells were treated with PMA at 50ng/ml final concentration in 2% FCS supplemented RPMI-1640 for 16 hours, followed by addition of Lipofectamine 2000 (complexes with 25 or 12.5 pmol of miR-185 specific antagomir (Exiqon) or scramble control (Exiqon) in serum free RPMI-1640 for 4 hours. Subsequently cells were supplemented with RPMI-1640 containing 2% FCS for 24 hours prior to cell lysis for total RNA extraction or total protein analysis.
Tsitoura E., Wells A.U., Karagiannis K., Lasithiotaki I., Vasarmidi E., Bibaki E., Koutoulaki C., Sato H., Spandidos D.A., Siafakas N.M., Sourvinos G, & Antoniou K.M. (2016). MiR-185/AKT and miR-29a/Collagen 1a pathways are activated in IPF BAL cells. Oncotarget, 7(46), 74569-74581.
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