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. 2022 Jul 4;13:931021. doi: 10.3389/fimmu.2022.931021

Figure 2.

Figure 2

TLR22-induced (Ca2+)mt flux instigates pro-apoptotic mtROS generation in A. hydrophila-infected HKM. HKM pre-incubated with YCG063, RR or transfected with sc-siRNA, tlr22-siRNA were infected with A. hydrophila and at 4 h p.i. (A) changes in mtROS levels were measured, and (B) changes in mtROS levels were visualized under fluorescence microscope. Vertical bars denote mean ± SEM (n=5). Fluorescence microscopic data is representative of three independent experiments. (C) HKM pre-incubated with RR or transfected with sc-siRNA, tlr22-siRNA were infected with A. hydrophila, and changes in (Ca2+)mt were measured using Rhod-2/AM at 1 h p.i. (D) HKM pre-incubated with YCG063, di-BPA, CsA, Mdivi-1 or transfected with sc-siRNA, tlr22-siRNA were infected with A. hydrophila and bacterial load were enumerated at 24 h p.i. Data represent mean ± SEM (n=5). Asterisk (*) signifies significant difference between the indicated group (*p < 0.05). HKM, uninfected HKM; HKM+B, HKM infected with A. hydrophila; HKM+YCG063+B, HKM+RR+B, HKM+di-BPA, HKM+CsA+B, HKM+Mdivi-1, HKM pre-incubated with YCG063, Ruthenium Red, di-BPA, CsA, Mdivi-1 were infected with A. hydrophila; HKM+sc-siRNA, sc-siRNA transfected HKM; HKM+sc-siRNA+B, HKM+tlr22-siRNA+B, sc-siRNA and tlr22-siRNA transfected HKM were infected with A. hydrophila.