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ATP assays were performed using ATP Assay Kit (MAK190, Sigma-Aldrich), according to the manufacturer’s instructions. Briefly, washed 1 × 106 Caco-2/LoVo cells were lysed in 100 μL of ATP assay buffer and deproteinized using a 10 kDa MWCO spin filter. Next, 80 μL of the samples was added into 50 μL of the reaction mix (ATP probe+ ATP converter+ developer mix) and incubated at room temperature for 30 minutes protected from light. ATP values were measured by the fluorescence (FLU, λex = 535/λem = 587 nm) in a microplate reader.
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