GSM7770306: Gal3-treatment, replicate 1, snRNAseq; Mus musculus; RNA-Seq
Frozen SDH tissue from C57BL/6J wild-type mice was washed in pre-cooled PBSE (PBS buffer containing 2 mM EGTA). GEXSCOPE® Nucleus Separation Solution (Singleron Biotechnologies) was used to isolate the single nucleus, referring to the manufacturer's product manual. The isolated nuclei were re-suspended in PBSE to 106 nuclei per 400 μl and then filtered through a 40 μm cell strainer. Finally, the nuclei were counted by Trypan blue. DAPI (1:1,000) (TermoFisher Scientific, D1306) was used to stain the nuclei enriched in PBSE, which were defined as DAPI-positive singlets. Single nucleus suspension (3×105-4×105 nuclei/ml) with PBS was loaded onto a microfluidic chip (GEXSCOPE® Single Nucleus RNA-seq Kit, Singleron Biotechnologies). Subsequently, the snRNA-seq libraries were constructed based on the manufacturer's instructions (Singleron Biotechnologies). The resulting snRNA-seq libraries were pooled for sequencing on the Illumina novaseq 6000 with 150 bp paired-end reads.