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To inactivate LS GLP-1R neurons, we injected AAV expressing double-floxed EGFP and TeNT (AAV-DIO-EGFP-2A-TeNT) into the bilateral LS of GLP-1R-ires-Cre mice to achieve specific expression of EGFP-2A-TeNT in LS GLP-1R neurons (Figure 3A and Supplemental Figure 6C)

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After incubation in 2 mL of 0.05% trypsin-EDTA (25300054, Thermo Fisher Scientific) for 5 min at room temperature, cells were transferred into 8 mL of culture medium, centrifuged for 1 min at 200 g , resuspended in fresh medium and re-seeded into a new plate for continuous culture or seeded into a specific plate at the desired cell density, depending on the experimental setup

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Gather the injection pen, new needle, and alcohol swab