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. 2021 Oct 29;12:741219. doi: 10.3389/fphar.2021.741219

FIGURE 3.

FIGURE 3

SC75741 inhibits TNF-α-induced aggregation formation and promotes aggregation clearance. (A). H4GT25 cells were pretreated with 1 μg/ml DOX for 24 h, treated with TNF-α (0, 1, 5, 10, and 20 ng/ml) for another 24 h, the GFP fluorescence was analyzed by fluorescence microscopy. (data represent mean ± SD; n = 6, **** p < 0.0001, ns, not significant, two-tailed t test). (B). H4GT25 cells were pretreated with 1 μg/ml DOX for 24 h, treated with TNF-α (0, 1, 5, 10, and 20 ng/ml) for another 24 h. Cell lysates were immunoblotted with indicated antibodies. The numbers under the blot represent the gray scale quantification (GFP/Tubulin, NF-κB/Tubulin, p-NF-κB/Tubulin). (C). H4GT25 cells were pretreated with 1 μg/ml DOX for 24 h, treated with TNF-α (20 ng/ml), JSH-23 (10 μM) or SC75741 (5 μM) for another 24 h. Cell lysates were immunoblotted with indicated antibodies. The numbers under the blot represent the gray scale quantification (GFP/Tubulin, NF-κB/Tubulin, p-NF-κB/Tubulin). (D). H4GT25 cells were pretreated with 1 μg/ml DOX for 24 h, treated with TNF-α (20 ng/ml), DMSO, JSH-23 (10 μM) or SC75741 (5 μM) for another 24 h, the GFP fluorescence was analyzed by fluorescence microscopy. (data represent mean ± SD; n = 4, **** p < 0.0001, *** p < 0.001, ns, not significant, two-tailed t test). (E). SH-SY5Y cells were transfected with GFP-SOD1 G93A for 24 h, treated with TNF-α (20 ng/ml), JSH-23 (10 μM) or SC75741 (5 μM) for another 24 h. Cell lysates were immunoblotted with indicated antibodies. The numbers under the blot represent the gray scale quantification (GFP/Tubulin, NF-κB/Tubulin, p-NF-κB/Tubulin). (F). SH-SY5Y cells were transfected with GFP-SOD1 G93A for 24 h, treated with TNF-α (20 ng/ml), JSH-23 (10 μM) or SC75741 (5 μM) for another 24 h, the GFP fluorescence was analyzed by fluorescence microscopy. (data represent mean ± SD; n = 6, **** p < 0.0001, * p < 0.05, two-tailed t test).