A promising drug candidate discovered for the prevention of type 1 diabetes helsinkiuni NatureComms
) to estimate the level of contamination. The Seurat analysis was then repeated with the adjusted counts with the following modifications. Cells with less than 5000 UMI counts or 1700 expressed genes were excluded. We also removed cells with unusually high level of mitochondrial reads . We assigned a cell cycle phase to all the cells using the default settings in CellCycleScoring-function and used this to remove biases due to cell cycle heterogeneity when scaling the data.
against the KEGG. ClusterProfiler was also used for geneset enrichment analysis against the same database, using fold change to rank the genes.Animal care and experiments were approved by National Animal Experiment Board in Finland . NOD-SCID-Gamma mice were obtained from SCANBUR and housed at Biomedicum Helsinki animal facility, on a 12 h light/dark cycle and fed standard chow ad libitum, 2016 Teklad global 16% protein rodent diets .
. At the end of S4, cells were dissociated into single cell suspension using TrypLE for 10 min at 37 °C and 5 × 10ells were electroporated with 2 µg of the plasmid using Neon Transfection system . Cells were then plated in AggreWell plates to form 3D aggregates for 24–48 h.For protein extraction, cells were washed with ice-cold PBS and lysed with Cell lysis buffer for 10 min on ice.
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