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TMH does not affect other inflammasome or inflammatory pathways. (A,B) LPS-primed J774A.1 cells were treated with TMH for 2 h, followed by stimulation with either dsDNA (1 μg/mL) (A) or flagellin (B) transfected using Lipofectamine 2000 for 3 h. (C) Following TMH treatment (2 h), nuclear and cytosolic fractions of LPS-primed J774A.1 cells were separated and assessed for NF-κB p65 translocation by immunoblotting. (D) HEK293FT cells transfected with the pGL4.32 luciferase reporter vector were treated with TNF-α (20 ng/mL) for 5 h, with or without TMH pre-treatment (2 h). Luciferase activity was measured using the Bright-Glo™ luciferase assay system (Promega). Data are presented as the mean ± SD (n = 3). Statistical analysis was performed using one-way ANOVA followed by Tukey’s multiple comparisons test n.s., not significant.

Journal: Frontiers in Pharmacology

Article Title: Thymelaea hirsuta (L.) Endl . extract attenuates NLRP3 inflammasome activation via modulation of ATPase activity

doi: 10.3389/fphar.2026.1781860

Figure Lengend Snippet: TMH does not affect other inflammasome or inflammatory pathways. (A,B) LPS-primed J774A.1 cells were treated with TMH for 2 h, followed by stimulation with either dsDNA (1 μg/mL) (A) or flagellin (B) transfected using Lipofectamine 2000 for 3 h. (C) Following TMH treatment (2 h), nuclear and cytosolic fractions of LPS-primed J774A.1 cells were separated and assessed for NF-κB p65 translocation by immunoblotting. (D) HEK293FT cells transfected with the pGL4.32 luciferase reporter vector were treated with TNF-α (20 ng/mL) for 5 h, with or without TMH pre-treatment (2 h). Luciferase activity was measured using the Bright-Glo™ luciferase assay system (Promega). Data are presented as the mean ± SD (n = 3). Statistical analysis was performed using one-way ANOVA followed by Tukey’s multiple comparisons test n.s., not significant.

Article Snippet: For inflammasome stimulation, ultrapure LPS (tlrl-ppglps), nigericin (tlrl-nig), adenosine triphosphate (ATP; tlrl-atpl), and flagellin (FLA-ST; tlrl-epstfla) were obtained from InvivoGen (United States).

Techniques: Transfection, Translocation Assay, Western Blot, Luciferase, Plasmid Preparation, Activity Assay