RNA-seq of human HEK-293T cells treated with a nutraceutical formula (solution-3), or vehicle-(saline solution)-treated controls
RNA-seq of human HEK-293T cells treated with a nutraceutical formula (solution-3), or vehicle-(saline solution)-treated controls
HEK-293T cells were treated with a nutraceutical formula containing propolis, Verbascum, thymus, long chain length polyphosphates, potassium sorbate and sodium benzoate (solution-3) at 0.01x concentration or with vehicle (saline solution) as the negative control for the treatment. After 24 hours, the cells were lysed with TRIzol RNA Isolation Reagent (1 ml, #15596018; Ambion, Thermo Fisher Scientific) and frozen at -80°C. HEK-293T cells were grown in a humidified 37°C incubator with 5% CO2. The cells were cultured in feeder-free conditions using Dulbecco's modified Eagle's medium (41966-029; Gibco) with 10% fetal bovine serum (10270-106; Gibco), 2 mM L-glutamine (25030-024; Gibco), and 1% penicil-lin/streptomycin (P0781; Sigma-Aldrich), with medium changed daily. Cells were dissociated with Trypsin-EDTA solution (T4049, Sigma-Aldrich) when the culture reached ~80% confluency. HEK-293T cells were plated (1000000 cells) and treated with a nutraceutical formula containing propolis, Verbascum, thymus, long chain length polyphosphates, potassium sorbate and sodium benzoate (solution-3) at 0.01x concentration or with vehicle (saline solution) as the negative control for the treatment. After 24 hours, the cells were lysed, and their RNA were extracted. Total RNA, isolated from HEK-293T cells by using TRIzol RNA Isolation Reagent (#15596018; Ambion, Thermo Fisher Scientific) was quantified in a NanoDrop™ One/OneC Microvolume UV-Vis Spectrophotometer (Thermo Scientific™), checked for purity and integrity. Libraries were prepared using the TruSeq® Stranded mRNA Library Kit according to the protocols recommended by the manufacturer. Library kit: TruSeq® Stranded mRNA