Illumina NovaSeq 6000 paired end sequencing; Effects of increased intracellular calcium and overnutrition on pancreatic beta cell transcriptome
Effects of increased intracellular calcium and overnutrition on pancreatic beta cell transcriptome
Ins2Apple/+ pancreatic islets were collected from mice at 8-9 weeks of age. Mice were segregated by sex, and the islets from 2-3 mice of the same sex and genotype were pooled for each sample. Islets were dispersed using Accumax and FACS purified using Apple fluorescence and DAPI viability dye, to collect only live pancreatic beta cells. Abcc8 KO and wild type mice containing Ins2Apple/+ allele were maintained in a C57BL/6J background. All mice used for experiments were fed either a standard chow diet or a 60% high fat diet for 5 week and were maintained on a 12-hour light/dark cycle. Total RNA was extracted from FACS-purified pancreatic beta cells using the Maxwell simplyRNA kit (Promega). RNA was amplified using the SMART-Seq v4 Ultra Low Input RNA Kit (Takara), and cDNA libraries were constructed using the Low Input Library Prep Kit (Takara).
Experimental Factor: sex
male
Experimental Factor: diet
normal
Experimental Factor: genotype
Abcc8-/-