On day 28 following PBOO operation, the animals were sacrificed and the bladders were removed guaranteeing the bladder weight. The bladder was sliced into longitudinal detrusor strips and divided into three sections for RT-qPCR, Western blot, and histological test. RT-qPCR was used to quantify the expression of SLC17A9 and ChAT. The methods were conducted as described in a previous article32 (link), using an EZ-press RNA Purification Kit, Color Reverse Transcription Kit, and 2*Color SYBR Green qPCR Master Mix (ROX2 plus), from EZBioscience, USA. Table 2 contains a list of primer pairs. Results were standardized using the expression of GAPDH.
Primers used for RT-qPCR.
| Gene | Primers (5′–3′) |
|---|---|
| SLC17A9—F | GCTTCCTCAAGGCTATGATCTT |
| SLC17A9—R | AGGTCCTGAATGTTGACTGAAA |
| ChAT-F | TGGCCATACCCAGGACACA |
| ChAT-R | TCCAAGACAAAGAACTGGTTGCA |
| GAPDH-F | TGAGCATCTCCCTCACAATTCC |
| GAPDH-R | TTTTTGAGGGTGCAGCGAAC |
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