“…Primary, wild type suture cells were isolated as previously described (Durham et al, 2019; Durham et al, 2016), plated at a density of 65,000 cells per well in 6 well plates and treated for 7 days with control media (DMEM, supplemented with 10% FBS, 1% penstrep, and 0.2% amphotericin) or media supplemented with pharmacological agents (levothyroxine (780 ng/ml), nicotine (25 ng/ml), or citalopram (250 ng/ml)) at levels mimicking circulating levels for 7 days (Durham et al, 2019; Durham et al, 2015). Subsequently, total RNA was isolated using the Qiagen RNEasy mini kit (Qiagen, Valenica, CA, USA) according to manufacturer’s protocol.…”