The material used in this study was Microcystin-leucine arginine (MC-LR) (ALX-350-012-M001; Enzo Life Sciences, NY, USA). To prepare the concentrate solution, 1 mg of MC-LR was suspended in 1000 μL of dimethyl sulfoxide (DMSO), yielding a concentration of 1 mM. To obtain 1, 10, 100, and 1000 nM of MC-LR for each experiment, the concentrate solution was diluted with the completed DMEM medium. These concentrations of MC-LR for treatment groups were selected based on microcystin-polluted surface waters, typically below 100 μg/L (equivalent to approximately 100.5 nM) [29 (link)], as well as many previous studies [23 (link),[30] (link), [31] (link), [32] (link)] that have utilized this concentration range.
The β-catenin inhibitor, MSAB (HY-120697; MedChem Express, NJ, USA) was first diluted with DMSO to prepare the stock solution. For the working solution, the inhibitor was further diluted with completed DMEM medium to achieve a 10 μM of MSAB. The inhibitor was treated to the cultured cell 24 h before MC-LR treatment.
The β-catenin inhibitor, MSAB (HY-120697; MedChem Express, NJ, USA) was first diluted with DMSO to prepare the stock solution. For the working solution, the inhibitor was further diluted with completed DMEM medium to achieve a 10 μM of MSAB. The inhibitor was treated to the cultured cell 24 h before MC-LR treatment.
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