Journal: iScience
Article Title: Haemolysins are essential to the pathogenicity of deep-sea Vibrio fluvialis
doi: 10.1016/j.isci.2024.109558
Figure Lengend Snippet: The cytotoxicity of δVFH (A–C) THP-1 cells were incubated with or without (Control) different concentrations of δVFH for 2 h and then subjected to microscopic observation (A), LDH release determination (B), and western blot using anti-Casp1, anti-GSDMD, or anti-β-actin (loading control) antibodies (C). The arrows in (A) indicate representative cells at different stages of death. Scale bar, 10 μm. (D) PBS or δVFH was incubated with wild type (WT) THP-1 cells or THP-1 cells defective in Casp1 (Casp1-KD) or NLRP3 (NLRP3-KD) for 2 h. The cells were subjected to LDH release determination (left) and western blot using anti-Casp1, anti-GSDMD, or anti-β-actin (loading control) antibodies (right). (E) PBS or δVFH was incubated with wild type (WT) THP-1 cells or THP-1 cells defective in ASC (ASC-KO) or GSDMD (GSDMD-KO) for 2 h. The cells were subjected to LDH release determination (left) and western blot (right) as aforementioned. (F–H) THP-1 cells were pretreated with different concentrations of the inhibitor MCC950 (F), Ac-YVAD-CMK (G), or NSA (H) for 1 h, and then treated with δVFH for 2 h. LDH release was then determined. (I and J) THP-1 cells were treated with or without KCl (0–100 mM) (I) or BAPTA-AM (0–40 μM) (J) for 45 min and then incubated with δVFH for 2 h. The cells were subjected to LDH release determination (left panels) and western blot using anti-Casp1, anti-GSDMD, or anti-β-actin (loading control) antibodies (right panels). The results of all histograms are the means of triplicate experiments and shown as means ± standard deviation (SD). ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001.
Article Snippet: Casp1-KD THP-1 cells , Invivogen , Cat# thp-dcasp1.
Techniques: Incubation, Control, Western Blot, Standard Deviation
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