Data files

Study of oligodendrocytes and microglia in aging in a tissue-specific manner (white and gray matter) using the 10X single-cell RNA-sequencing platform. We also analyzed Rag1-deficient mice to study how the lack of mature T cells affect the age-dependent glial alterations that occur in the white matter.

Study of microglia in aging and in a tissue specific manner (white and grey matter) using Smart-seq2 and 10x scRNA-seq technologies. We also analysed, 5xFAD mice (AD model) with Smart-seq2, and Apoe-KO with 10x transcriptomics. Additionally we studied the impact of Actinomycin-D during tissue dissociation on gene expression in microglia using Smart-seq2.

Grey and white matter (corpus callosum) were dissected from the brain of 4 months and 21 months old wild type mice, single cell suspension was isolated using Neural Tissue Dissection kit (myelin was removed on percoll gradient) then microglia cells were isolated using CD11b microbeads by Magnetic separation with MS columns (MACS technology).The cell pellets are resuspended in the RLT buffer (100ul-150ul), snap frozen in liquid nitrogen and stored in -80 freezer. RNA was extracted and transcriptome ...

RNA-seq of sorted cells from brain of Cortical MS mouse treated with vehicle or CSF1R inhibitor

To study the effect of the Grk2 or Ets1 KOs in rat and human CD4+ T cells, in vivo or in vitro respectively, we generated KO CD4+ T cell lines by CRISPR-RNP nucleofection (with an gRNA targeting the gene locus) or control (with a non-targeting gRNA) 3' bulk RNAseq was conducted on CD4+ T-MBP rat cells isolated form spleen or spinal cord parenchyma three days after co-transfer of control and KO cells into the animal. Cells were FACS sorted based on their fluorescent protein reporters, BFP for ...

To identify genes driving encephalitogenic CD4+ T cell migration into the CNS, we performed a genome-wide CRISPR screen and a subsequent validation screen. For the genome-wide CRISPR screen, up to 4 sgRNA per gene and 800 non targeted controls were included, for a total of 87690 individual sgRNAs, and for the validation screen, up to 6 sgRNA per gene and 241 non targeted controls for a total of 12000 individual sgRNAs. Comparison of sgRNA distribution by pairwise comparisons across blood, spleen, ...

To study the regulation of candidate genes from our study in human cells, we analyzed CD4+ T cells from blood and CSF of MA patients and age and sex matched idiopathic intracranial hypertension controls We analyzed 40845 cells in control blood, 807 cells in control CSF, 29749 cells in MS blood and 15768 cells in MS CSF. Analysis of overlapping TCR sequences in blood and CSF CD4+ T cells of the same transcript, transcriptomic characterization of the cell clusters and analysis of the behaviour of ...