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Cells were washed and imaged in RPMI-1640 without phenol red (32404014, Thermo Fisher Scientific), while islets were washed and imaged in 6 mM glucose KRBH buffer using a Nikon spinning disk field scanning confocal microscope with a 60X oil objective at 37C

The titration schedule itself (0.25 0.5 1.0 1.7 2.4 mg at ~4-week steps) is not a research artifact

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Between the suppressed appetite, the loss of thirst cues, and the occasional waves of nausea or fatigue, water alone wasnt cutting it

This allows for meticulous quality control at every step

Oregon R + was used as the wild type control line for all experiments