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For starved conditions, HeLa wildtype and knockout cells were grown to near confluency in Greiner Bio-One 10-well glass-bottom dishes, starved in EBSS, loaded with Lysotracker red, and imaged live on a NikonCSU W1 spinning disc microscope equipped with environmental control (humidity, 37 C, and 5% CO 2 ) using a 60x oil objective

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Triple agonists add glucagon receptor activation, which: Increases resting energy expenditure by 8% to 12% (Jall et al., Diabetes Care 2024) Promotes hepatic fat oxidation and reduces steatosis May offset some of the lean mass loss seen with GLP-1 monotherapy (early signal, not yet confirmed in long-term trials) The weight loss ceiling for retatrutide appears to be 24% to 26% based on phase 3 data, though long-term real-world outcomes are not yet available