Journal: The Biochemical journal

Article Title: FoxA2 and RNA Pol II Mediate Human Islet Amyloid Polypeptide Turnover in ER-stressed Pancreatic β-cells

doi: 10.1042/BCJ20200984

Figure Lengend Snippet: RIN-m5f cells were transduced with empty vector (EV), hIAPP or rIAPP encoding lentivirus particles for 48h. (A) Immuno-confocal microscopy optical section (1μm-Z plane) of RIN-m5f cells transduced with empty vector (EV) and hIAPP encoding lentivirus. Cells were co-stained with hIAPP-specific IAPP monoclonal antibody (red) and DAPI (blue). (B) Representative single (1μm-Z plane) fluorescence confocal sections of hIAPP intracellular accumulation sites in hIAPP lentivirus-transduced cells. Arrows denote hIAPP’s nuclear locations in micrographs. Bars, 5μm.

Article Snippet: Lentiviral particle production and lentivirus-mediated transduction Flag-tagged ORF sequences for pre-pro-hIAPP and rIAPP, flanked with restriction enzyme sites (5’ SgfI and 3’ Mlu1) were synthesized using gBlock gene synthesis (Integrated DNA Technology) and cloned into a third-generation lentivirus destination vector (pLenti-C-Myc-DDK-IRES-Puro, Origene, cat# PS100069).

Techniques: Transduction, Plasmid Preparation, Confocal Microscopy, Staining, Fluorescence