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For the H&E study, the rats were sacrificed by cervical dislocation without prior anesthesia, and their hearts, livers, kidneys, spleens, and intestines were immediately dissected, stored at − 80 °C, and snap-frozen in isopentane on dry ice until further processed.
Bovine bladder samples collected as above described were stored at −80°C until further processed.
The remaining part was frozen in isopentane pre-cooled in liquid nitrogen and stored at −80°C until further processed for immunohistochemistry.
The organoids were then frozen in 6% dimethyl sulfoxide (DMSO, Fluka, Milan, Italy) containing medium and stored at −80°C until further processed.
The pelleted nuclei were dissolved in 1 mL 1×PBS, centrifuged for 5 min at 14,000×g, and the nuclei pellets stored at −80°C until further processed.
Tumor biopsies were frozen in liquid nitrogen until further processed.
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The sections were stored in the dark at 8°C until being further processed.
Crude homogenates were aspirated 5 10-times through a 0.8 mm syringe needle and stored at −80°C until being further processed.
Samples were stored at -80 °C until they were further processed for metabolite analysis.
DNA was stored at 4°C until the samples were further processed.
Total RNA was further processed in RT-PCR or stored at −80°C until its use.
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