RNA-seq of kidneys from mice treated for eight weeks with the glucagon analogue, NNC9204-0043, compared to mice treated with PBS
RNA-seq of kidneys from mice treated for eight weeks with the glucagon analogue, NNC9204-0043, compared to mice treated with PBS
The mice were fasted overnight (11 hours) prior to liver isolation, and livers were kept at -80°C until analysis. C57Bl/6JRj mice (8 weeks of age) received either a long acting glucagon analogue (NNC9204-0043; terminal half-life of 5-6 hours, 1.5 nmol/kg twice daily) or PBS+1% BSA (twice daily). Drugs were administered over a period of 8 weeks, by subcutaneous injection. The initial dose of the glucagon analogue of 3 nmol/kg was reduced to 1.5 nmol/kg after 16 days of treatment, due to a weight decrease of >20% of one of the mice (the mouse was terminated and excluded from the study). Raw sequencing data were processed using bcl2fastq software version 2.20.0 (Illumina). Total DNA/RNA was purified by an AllPrep DNA/RNA Minikit (catalog no. 80204; QIAgen, Hilden, Germany) according to manufacturer's instructions and quality tested using a 2100 Bioanalyzer instrument (Agilent Genomics, Santa Clara, California, USA). The extracted RNA was DNase treated with RNeasy Minikit (catalog no. 74106; QIAgen, Hilden, Germany) and RNase-Free DNase set (catalog no. 79254; QIAgen, Hilden, Germany) according to manufacturer's protocol. All the above-mentioned processes were performed on a QIACube machine (QIAgen, Hilden, Germany). Library preparation was done using TruSeq Stranded Total RNA Library Prep Gold (catalog no. 20020599; Illumina, San Diego, California, USA) and IDT for Illumina-TruSeq RNA UD Indexes (cat 20022371; Illumina, San Diego, California, USA) according to the manufacturer's instructions