Single-cell gene expression profiles of pancreatic islets from two β-cell specific knockout mouse models (βZKO-Mip and βZKO-Rip), along with RIP-Cre and MIP-Cre mice, were analyzed after being fed a high-fat diet (HFD).

The islet is composed of diverse cell types that work together to regulate its endocrine function. To investigate this, we performed comparative single-cell RNA sequencing (scRNA-seq) on islets from 16-week-old βZKO-Mip, βZKO-Rip, RIP-Cre, and MIP-Cre mice, all of which were fed a high-fat diet (HFD) from 12 to 16 weeks. Sequencing was carried out using the Chromium Next GEM Single Cell 3' Reagent Kits. Each islet sample was derived from at least three mice with the same genotype. Briefly, two β-cell-specific knockout mouse models (βZKO-Mip and βZKO-Rip) were generated by crossing C57BL/6N mice carrying floxed exon 15 (Zzef1f/f) with MIP-Cre or RIP-Cre mice. Each islet sample was derived from at least three mice with the same genotype. The purified islets were then treated with 2 mL of 0.05% TryLE at 37°C for 4 minutes, followed by gentle pipetting (~20 times). The resulting cell suspension was stained with 0.4% trypan blue to assess cell viability under a microscope. Cells with viability greater than 85% were sorted and used for 3’ digital gene expression profiling, with approximately 10,000 individual cells per sample, using the 10x Chromium Single-Cell 3’ Reagent Kit (10x Genomics, USA) according to the manufacturer’s instructions.