Nextflex poly a kit

We performed RNA-sequencing as described previously [41 (link)]. Total RNA was isolated from renal tissue as described in the Method of qRT-PCR. The concentration, quality, and integrity of RNA were checked by using Nanodrop and Bioanalyzer (RNA 6000 Nano Kit, Agilent Technologies, Santa Clara, CA). Five micrograms of high-quality tissue RNA were used to obtain poly A–enriched RNA by using the NEXTflex Poly(A) Kit (Bioo Scientific, Austin, TX). Next, the mRNA concentration was measured by using Qubit (HS RNA assay kit, Agilent Technologies), and 50 ng was used to prepare mRNA libraries by using the NEXTflex Rapid Directional mRNA-Seq Kit (Bioo Scientific). The library concentration and size were measured by using the KAPA Library Quantification Kit (Kapa Biosystem, Wilmington, MA) and Bioanalyzer, respectively. The libraries were then subjected to 100 bp paired-end next-generation sequencing (Illumina, SanDiego, CA). To visualize the distribution of reads on the NLRP3 signaling genes, BedGraph files were generated from RNA-sequencing data by using HemI 1.0-Illustrator software (The CUCKOO Workgroup, School of Life Science and Technology, Huazhong University of Science and Technology, China).