Data files

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 ...

Sample preparation for proteomics analysis was performed as described previously with slight modifications. Briefly, for mouse samples, SDC lysis buffer (2% SDC, 100 mM Tris-HCl pH 8.5) was used to lyse the cell pellets at 95°C for 45 min at 600 rpm in a thermoshaker. For human samples which were fixed in PFA, prior to the SDC lysis buffer step, the samples were first resuspended in 6% SDS buffer, heat denatured, sonicated and then precipitated using 80% acetone overnight in -20°C. Next day, these ...

After respective treatment, cells were washed 2 times with ice-cold DPBS, scraped in PBS and pellets either processed immediately or stored at -80°C. Cells were lysed in RIPA buffer for 30 min on ice. After clearance by centrifugation at 20,000 × g for 10 min at 4 °C, protein concentrations were adjusted using BCA assay. Cleared and adjusted supernatants were incubated overnight on an overhead rotator with preequilibrated streptavidin agarose (Sigma). Next day, beads were washed 2 times with RIPA ...