title: GSM8035276: 7dpi_cKO; Mus musculus; RNA-Seq
library_construction_protocol: Immediately following euthanasia, mice were transcardially perfused (10mL DPBS), and skullcaps with lesions were removed and placed in iMED+ (15mM HEPES and 0.6% glucose in HBSS with phenol red). Lesions were dissected; tissue from male and female mice within experimental conditions were combined. Tissue was minced for 10 minutes in 100mL of TST buffer (146 mM NaCl, 10 mM Tris-HCl pH 7.5, 1 mM CaCl2, 21 mM MgCl2, 0.03% Tween-20, and 0.01% BSA in nuclease-free water), followed by filtration (40mm), and washing with TST and ST buffer (146 mM NaCl, 10 mM Tris-HCl pH 7.5, 1 mM CaCl2, and 21 mM MgCl2). Samples were centrifuged (500g 10 minutes), resuspended (PBS/1% BSA/0.2UmL Protector RNAse inhibitor), filtered (35mm), re-centrifuged (500g 5 minutes), and resuspended in FANS buffer with 0.2U/mL RNase inhibitor and 2mg/mL DAPI. Nuclei were subsequently sorted (FSC/SSC/DAPI). All samples were centrifuged (500g 15 minutes) and resuspended to a maximum of 16,500 nuclei in 45uL. Up to 1.6 x 10^4 nuclei were loaded onto the Chromium Controller (10X Genomics). Chromium Single Cell 3' v3.1 reagents were used for library preparation according to the manufacturer's protocol.