GSM5503749: MUC26021; Mus musculus; RNA-Seq
Anesthetized mice underwent an intravascular staining procedure [20] with 0,75 µg of TotalSeq™-A0096 anti-mouse CD45 Antibody (clone 30-F11, Biolegend) in 300 µl PBS, 3-10 minutes before CSF aspiration from the cisterna magna, to be able to discriminate CSF from blood leukocytes. After thorough perfusion with PBS the spinal cord was dissected, and meninges were detached from the spinal cord using fine forceps. Meninges were passed through a cell strainer to collect mELT cells. Fc receptors were blocked and samples were incubatedwith DNA barcoded TotalSeq antibodies against CD4 (TotalSeq-C0001 anti-mouse CD4, clone RM4-5, Biolegend), CD8a (TotalSeq-C0002 anti-mouse CD8a , clone 53-6.7, Biolegend), CD19 (TotalSeq-C0093 anti-mouse CD19, clone 6D5, Biolegend), Ly6G (TotalSeq-C0015 anti-mouse Ly6G , clone 1A8, Biolegend), and NK1.1 (TotalSeq-C0118 anti-mouse NK-1.1, clone PK136, Biolegend), and one of 2  TotalSeq Hashtag antibodies (TotalSeq™-C0301 anti-mouse Hashtag 1 Antibody, clones M1/42; 30-F11, Biolegend or TotalSeq™-C0302 anti-mouse Hashtag 2 Antibody, clones M1/42; 30-F11, Biolegend), to allow pooling of 2 samples per experiment. 10x Genomics Chromium Next GEM Single Cell V(D)J Reagent Kit (v1.1) 10x Genomics single cell sequencing, with feature barcoding