GSM9111312: Lung 1; Mus musculus; RNA-Seq
Before tissue extraction, lungs were perfused by injecting PBS through the right ventricle. The extracted pieces were frozen in liquid nitrogen. For kidney samples, half of a frozen mouse kidney was minced on dry ice and homogenized in 5 ml of ice-cold homogenization buffer until no visible chunks remained. Samples from 3 mice were pooled. A 200 µl aliquot of the pooled homogenate was mixed with 400 µl of cold lysis buffer from the Minute™ Single Nucleus Isolation Kit (Invent Biotechnologies) and processed according to the manufacturer's protocol. Nuclei were filtered, centrifuged, washed, and resuspended in PBS containing 1% BSA and 0.2 U/µl RNase inhibitor. Nuclei concentration was measured with Trypan Blue and adjusted to 700–1200 nuclei/µl. For lung samples, the protocol was similar, but only ~50 mg of frozen lung tissue was used for homogenization. The single-nucleus libraries were prepared with Single Cell 3' v4 (polyA) chemistry  (10x Genomics).