Qscript synthesis kit

Total RNA samples were isolated using the GeneElute kit, following the manufacturer’s protocol (Sigma-Aldrich). cDNA synthesis was carried out using the qScript synthesis kit, following the manufacturer’s protocol (Quantabio). mRNA expression was measured by quantitative (Q)-PCR using SYBR Green Mastermix (Eurogentec Ltd.) and the DNA Engine Opticon 2 system (BioRad). Q-PCR primer sequences were as follows: CHCHD4_F 5′-GAGCTGAGGAAGGGAAGGAT-3′; CHCHD4_R 5′-AATCCATGCTCCTCGTATGG-3′; KRT18_F 5′-TAGATGCCCCCAAATCTCAG-3′; KRT18_R 5′-CACTGTGGTGCTCTCCTCAA-3′; CDH2_F 5′-AGGATCAACCCCATACACCA-3′; CDH2_R 5′-TGGTTTGACCACGGTGACTA-3′; VIM_F 5′-GAGAACTTTGCCGTTGAAGC-3′; VIM_R 5′-TCCAGCAGCTTCCTGTAGGT-3′; NDI1_F 5'-AGTCAGATTCGCTTCCACCA-3'; NDI1_R 5'-CCCAGTATCAGCACGTTTGG-3';  ACTB_F 5′-CCCAGAGCAAGAGAGG-3′; ACTB_R 5′-GTCCAGACGCAGGATG-3′

Total RNA was extracted from frozen liver (~ 20 mg) using a RNeasy Plus Universal Mini Kit (QIAGEN), as per manufacturer’s instructions. The concentration of eluted RNA was measured using a Nanodrop DN-1000 spectrophotometer. cDNA synthesis was carried out using the qScript synthesis kit, following the manufacturer’s protocol (Quantabio). mRNA expression was measured by quantitative (Q)-PCR using SYBR Green Mastermix (Eurogentec Ltd.) and the DNA Engine Opticon 2 system (BioRad). Primers were obtained from QuantiTech Primer Assay (QIAGEN) and product details are as follows: Rn_Hk2_1 QT00190764, Rn_Pfkl_1 QT00175651, Rn_Ldha_2 QT02336243, Rn_Higd1a_1 QT00372428, Rn_Stoml2_1 QT01571724 , Rn_Slc25a11_1 QT01082914 , Rn_Actb_1 QT00193473. In the instance of Cox7a2l, QuantiFast SYBR Green PCR kit (QIAGEN) and QuantStudio 1 Real-Time PCR System (Applied Biosystems, Thermo Fisher) were used. The primer was obtained from QuantiTect Primer Assays (QIAGEN) with the following product details: Rn_Cox7a2l_1_SG. In all cases, transcript levels were normalised to levels of Actb and fold change determined using the 2^−ΔΔCT method, with expression in vehicle/normoxic animals normalised to 1.

Total RNA samples were isolated using the GeneElute kit (#RTN350), following the manufacturer’s protocol (Sigma-Aldrich). cDNA synthesis was carried out using the qScript synthesis kit (#95048-100), following the manufacturer’s protocol (Quantabio). mRNA expression was measured by quantitative (Q)-PCR using SYBR Green Mastermix (#RT-SY2X-NRWOU+B, Eurogentec Ltd.) and the DNA Engine Opticon 2 system (BioRad). The Q-PCR primer sequences are included in Additional file 2.