Quantifying Tnfxyd8 mRNA Expression

The methods used herein were modified from those described by Yang et al. (2013 (link)). To determine the abundance of Tnfxyd8 mRNA, total RNA was extracted from the whole kidney and purified using the RNA-Bee isolation kit (Tel-Test, Friendwood, TX, USA) and RNAspin Mini kit (GE Health Care, Piscataway, NJ, USA), respectively, following the manufacturer's instructions. RNA integrity was verified by 0.8% agarose gel electrophoresis. Extracted RNA samples were stored at −80°C after isolation. For reverse transcription, first-strand cDNA was synthesized using SuperScript™ Reverse Transcriptase (Invitrogen, Carlsbad, CA, USA) following the manufacturer's instructions. The cDNA products were stored at −20°C until analysis via polymerase chain reaction (PCR).