RNA-seq of human Caco-2 cells treated with a nutraceutical formula (solution-3), or vehicle-(saline solution)- control, and infected with SARS-CoV-2 (EG.5)
RNA-seq of human Caco-2 cells treated with a nutraceutical formula (solution-3), or vehicle-(saline solution)- control, and infected with SARS-CoV-2 (EG.5)
Caco-2 cells (ATCC, Middlesex, UK; accession number: HTB-37 L) were plated (500000 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 1 hour, the treated cells were infected with SARS-CoV-2 viral particles EG.5 (MOI, 3). After 48 hours, the infected cells were were lysed with TRIzol RNA Isolation Reagent (0.5 ml, #15596018; Ambion, Thermo Fisher Scientific) and frozen at -80°C. Caco-2 (ATCC, Middlesex, UK; accession number: HTB-37 L) 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 dis-sociated with Trypsin-EDTA solution (T4049, Sigma-Aldrich) when the culture reached ~80% confluency. Caco-2 cells (ATCC, Middlesex, UK; accession number: HTB-37 L) were plated (500000 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 1 hour, the treated cells were infected with SARS-CoV-2 viral particles EG.5 (MOI, 3). After 48 hours, the infected cells were were lysed, and their RNA were extracted. Total RNA, isolated from Caco-2 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