DRG Gene Expression Analysis in Asic1b Mice

The DRG samples of the lumbar parts were collected from 16-week-old female mice with Asic1b^+/+ or Asic1b^–/– genotypes. RNA extraction of the DRG samples was conducted with RNeasy Mini Kit (Qiagen) with on column DNase digestion (Qiagen), based on the manufacturer’s protocol. The RNA concentration and quality were measured by NanoDrop 1000 (Thermo Scientific). The cDNA reverse transcribed from RNA was prepared using with the iScript cDNA synthesis kit (Bio-Rad). Gene expression of DRG was measured using FastStart Universal SYBR Green Master with Rox (Roche) and analyzed using ABI 7500 Real Time PCR System (Life Technologies). Gene expression was normalized to gapdh mRNA. Data are presented as fold-change of gene expression in each group relative to the WT group of each gene. The primers of ASIC1a, ASIC1b, ASIC2a, ASIC2b, ASIC3, ASIC4, and gapdh were adapted from the paper of Wu et al. (2019) (link).