First weigh out 0.05 g cholesterol (Mw = 386.65 g/mol) and 0.45 g standard zebrafish larval food (SP; Sparos Lda; Zebrafeed <100 μm) and place in a 5 mL glass beaker to create a 10% w/w cholesterol-enriched diet, referred here as HCD. Components are homogenized and mixed by adding 1.5 mL diethyl ether that allows cholesterol solubilization. In another glass beaker, 1.5 mL diethyl ether is added to 0.5 g of SP to serve as a control diet (SPE; SP with diethyl ether). Diets are left overnight in the hood for the diethyl ether to evaporate completely and grounded up into fine particles using a pestle the following day.

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