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HeLa WT and CTS DBLZ-deficient as well as SH-SY5Y WT and CTS DBL-deficient cell clones (each n=6) were seeded 24 h after cell pellet collection. Cells were washed in ice cold PBS and harvested with a cell scraper. A modified RIPA lysis buffer (1% Triton X-100, 0.5% sodium deoxycholate, 0.1% SDS, 150 mM NaCl, 5 mM EDTA, 50 mM TrisHCl pH 8) was used to lyse the cells one ice for 30 min. The supernatant was collected after centrifugation at 16.000 rpm for 10 min at 4°C. 10 μl were used to determine ...

CD8+ T cells from PBMCs of the MS twin cohort (n = 12; 6 healthy twins, 6 twins with SCNI, and 12 twins with MS; 193,771 cells) and the validation cohort (n = 17; 5 individuals with IIH and 12 individuals with MS; 89,859 cells) were isolated via FACS and processed using the 10X manufacturer's protocol to generate single-cell transcriptome and TCR libraries. The corresponding TCR sequences are included within the metadata of the uploaded objects. (Author states that fastq raw data files from humans ...

We treated male Oxt-ires-Cre;RiboTag mice with either vehicle or CCK, collected hypothalami 2h post injection, pooled 2-3 tissues per sample, and affinity purified conditionally HA-tagged ribosomes (incl translating mRNA) specifically from oxytocin neurons. We then performed gene expression profiling analysis using data obtained from RNA-seq of immunoprecipitates versus inputs (n=4 per group) from standard chow diet fed mice and inputs only from high-fat/high-sugar diet fed mice (n=4 per group). ...

Hypothalamic oxytocin neurons from Oxt-ires-Cre;CAG-Sun1sfGFP mice were isolated by Fluorescence-activated nuclei sorting (FANS) according to the presence or absence of sfGFP signal and analyzed using snRNAseq2.

ARC tissue derived from C57BL/6JRj mice fed with SC diet (control), 5 days of HFHS (58%) diet, and 15 days of HFHS (58%) diet was dissociated by enzymatic digestion into single cells, which were then analyzed by scRNA-Seq. Per each experimental group, the ARCs from 6 animals were pulled into one sample.

C57BL/6JRj mice were exposed to a SC or a HFHS (58%) diet at 8-9 weeks of age for 4 months. Afterwards, hypothalami, hippocampi, and half cortices derived from 2 animals were isolated and pooled together in distinct tubes. A number of 5-7 replicates per each group was used. ACSA2+ astrocytes were isolated from each sample by magnetic-activated cell sorting (MACS), and further processed for RNA-Sequencing analysis.

A single-cell RNA sequencing experiment was conducted using 10x Genomics technology on samples from three distinct groups, which included six bones, meninges, and the brain. The study comprised three groups: those that underwent MCAo surgery, those that underwent sham surgery, and a group of naive animals. Each sample consisted of cells pooled from three animals, with a total of 32 samples utilized. The MCAo surgery group contributed 2 samples, each consisting of cells pooled from 3 different ...

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