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Fig. 2 Figure 2. Glucose uptake assay protocol. 2a. Equation detailing the reactions involved in a glucose uptake assay. 2b. Experimental setup of a glucose assay performed on 96 hpf zebrafish larvae exposed to different concentrations of caffeine. Larvae are injected with 2-deoxyglucose into the yolk (yellow area) and recovered for 30 min. Larvae are transferred to individual wells and lysed with acid detergent to stop further glucose uptake, destroy any native NADPH, and homogenise the sample. Neutralisation and detection buffer are added, and samples are incubated while the luminescent signal is generated. After 30 min, luminescence is measured on a plate reader.