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Islets were then washed with the Cholesterol Detection wash buffer, incubated for a minimum of 1 hour in Filipin III diluted in the Cholesterol Detection Assay buffer, and washed again in wash buffer before immediate imaging on a Zeiss LSM-780 inverted confocal laser-scanning microscope from the Facility for Imaging by Light Microscopy (FILM) at Imperial College London, with a 63X objective and a laser excitation wavelength of 340-380 nm and emission of 385-470 nm

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The HLCS gene is located on chromosome 21q22.13 and is composed of 19 exons that generate eight alternatively spliced mRNAs, seven of which encode the same 726 amino acid protein (isoform 1) and one which encodes a 873 amino acid protein (isoform 2)