Quantstudio 7 real time cycler

Prior to extraction of RNA using a RNeasy spin column (Qiagen), blood from the posterior vena cava was treated with red blood cell lysis buffer and lung tissues were treated with RNAlater solution (Ambion). All samples were DNase I treated (Qiagen) on the column during purification. For examination of SA100A8 transcripts, qRT-PCR analyses were performed using a SuperScript III One-Step RT-PCR kit (Thermo Fisher Scientific) on a QuantStudio 7 Real-Time Cycler (Thermo Fisher Scientific). Transcription levels of genes analysed were normalized to those obtained for ACTB. Data represent the mean (± S.E.M.) of two independent experiments with treatment cohorts comprised of at least six mice. The examination of mouse immune markers by qPCR was conducted using the TaqMan Array Mouse Immune Panel (Thermo Fisher Scientific) on a QuantStudio 7 Real-Time Cycler (Thermo Fisher). The TaqMan Arrays analyses were performed in triplicate, with each sample representing RNA pools from at least three mice. The results were analysed using the ThermoFisher Cloud Software. For the TaqMan Arrays analyses, data represent the mean (± S.E.M.), with statistical analyses performed using a one-way ANOVA.