β-catenin triggers ROS-induced-mtROS generation and downregulates sod2 mRNA expression. ZKM pre-treated with or without β-catenin inhibitors (JW67 and MSAB), NOX inhibitors (Apo and DPI), and mtROS inhibitor (YCG063) for 1 h were infected with A. hydrophila, and at 12 h p.i., (A) changes in mtROS levels were measured with fluorimetry, and (B) changes in mtROS levels were visualized under the fluorescence microscope. Fluorescence microscopic data are representative of three independent experiments. Ant A was used as a positive control. Additionally, ZKM were incubated with canonical Wnt/β-catenin pathway activators (LiCl and Laduviglusib), and changes in mtROS levels were measured with both fluorimetry and fluorescence microscopy. (C) ZKM pre-treated with or without the β-catenin inhibitor (JW67) were infected with A. hydrophila and sod2 mRNA expression was analyzed with RT-qPCR at 12 h p.i. Vertical bars denote the mean ± SEM (n = 3). Asterisk (*) denotes a significant difference between the indicated groups (* p < 0.05). “+B” mentioned in the X-axis and microscopy images represents “+A. hydrophila”.
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