library_construction_protocol: Sn protocol: the frozen tissue was homogenized in nuclei lysis buffer (NLB) containing 250mM sucrose, 10mM Tris-HCl, 3 mM MgAc2, 0.1mM EDTA, 0.1% Triton X-100 (Sigma-Aldrich, USA) and 0.2 U/μl RNase Inhibitor (Takara, Japan). Tissues were homogenized using a tissue grinder for 15-30 strokes at 2-8°C until they were lysed uniformly. Sucrose concentration was adjusted to purify the nucleus better. sc-RNA Seq sample concentration was adjusted to ~1000 nuclei/μl.single nucleus were collected using  the 10X Genomics Chromium Controller Instrument and Chromium Single Cell 3' V3.1 Reagent Kits (10X Genomics, Pleasanton, USA) . RNA libraries were prepared for sequencing using standard Illumina protocols