Nanodrop nd 1000 uv vis spectrophotometry

HUVECs were treated with 10 μmol/L HT for 1 h and then stimulated with IL-1β (0 or 5 ng/mL) for 3 h, after which the cells were collected and total RNA extracted using the Qiagen RNeasy kit (Qiagen, Milan, Italy) following the manufacturer’s instructions. The total RNA was stored at −80 °C until use. We determined the concentration and purity of RNA by NanoDrop ND-1000 UV-Vis Spectrophotometry (NanoDrop Technologies, Wilmington, DE, USA) and the integrity of RNA by using the Agilent 2100 Bioanalyzer (Agilent Technologies Inc., Santa Clara, CA, USA). The same RNAs were used for microarray and quantitative reverse-transcription polymerase chain reaction (qRT-PCR) analysis. In addition, we performed qRT-PCR analyses on different HUVEC samples to further confirm microarray data results. No variation in the total RNA yield was observed under the different experimental conditions tested (data not shown).

HUVECs were preincubated with 50 μmol/L DHA for 48 h, followed by stimulation with 5 ng/mL interleukin (IL)-1β, for additional 0–3 h, after which time cells were collected and total RNA extracted using the Qiagen RNeasy kit (Qiagen, Milan, Italy) according to manufacturer’s instructions. Concentration and purity of RNA was determined by NanoDrop ND-1000 UV-Vis Spectrophotometry (NanoDrop Technologies, Wilmington, DE, USA). No variation in the total RNA yield was observed under the different experimental conditions tested (data not shown).