, 5 glucose. To assess ENaC induced current, 50 µM amiloride was added to the luminal perfusate. ENaC activity was estimated from the effect of ENaC blocker amiloride on the transepithelial potential difference and current, which reflects the Nareabsorption from the colon epithelial layer. Transepithelial potential difference was determined continuously and transepithelial resistance estimated from the voltage deflections elicited by imposing rectangular test currents of 1 µA and 1.
FASTQ were generated using fastp and RNA-seq data quality assessed to identify potential issues with sequencing cycles, low average quality, adaptor contamination, or repetitive sequences from PCR amplification. Reads were aligned using STAR against a custom-built genome composed of the Ensembl rattus norvegicus mRatBN7.2. Alignment quality was analyzed using MappingQC and visually inspected in the Integrative Genome Viewer . Normalized read counts for all genes were obtained using edgeR .
The following primary antibodies were used for immunocytochemistry: mouse anti-α-Synuclein , rabbit anti-β-III tubulin . Primary antibodies were detected using the fluorescent secondary antibodies goat anti-rabbit Alexa 488 and goat anti-mouse Alexa 546 . Nuclear staining was carried out with DAPI solution . A Zeiss Axio Imager.Z1 fluorescence microscope with integrated Apotome module was used for microscopic evaluation and structured illumination imaging.