Journal: Redox Biology
Article Title: Lactate-driven ATP6V1B2 lactylation triggers asthmatic inflammation by linking lysosomal dysfunction to mitochondrial ROS-dependent pyroptosis
doi: 10.1016/j.redox.2026.104059
Figure Lengend Snippet: Effect of inhibiting Cathepsin B on mitochondrial dysfunction and tBid-Bax pathway. (A) Fluorescence imaging of mitochondrial membrane potential (JC-1) after administration of Cathepsin B inhibitor (Ca-074-Me). Scale bar: 100 μm. The ratio of red/green fluorescence intensity was quantified. (B) MitoSOX Red staining detects mitochondrial ROS. Scale bar: 100 μm. Quantification of MitoSOX Red fluorescence intensity is shown. (C) TEM images of mitochondrial ultrastructure. Scale bar: 2.0 μm. (D) Immunoblot analysis of Cathepsin B distribution in subcellular fractions (lysosome, cytosol, mitochondria). (E) Immunofluorescence staining of Cathepsin B (green) and LAMP1 (red). Scale bar: 100 μm. (F) Immunofluorescence staining of Cathepsin B (green) and MitoTracker (red). Scale bar: 40 μm. (G) Immunoblot analysis of Bid, tBid, XIAP, Bcl-2, Bcl-xL, and Bax protein expression. Data are expressed as Mean ± SEM. (n = 3 independent experiments) ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001, ∗∗∗∗P < 0.0001; ns, not significant.
Article Snippet: In mechanistic studies, cells were pre-incubated with the following specific inhibitors for 2 h before stimulation: Ferrostatin-1, Necrosulfonamide, Z-VAD-FMK (10 μM), Z-IETD-FMK (10 μM), Z-DEVD-FMK (10 μM), Bafilomycin A1 (100 nM), Chloroquine (10 μM), 3-Methyladenine (3-MA, 3 mM), Cathepsin B inhibitor CA-074 Methyl Ester (Cat# HY-100350, MCE), BID inhibitor BI-6C9 (Cat# HY-103661, MCE), and calcium chelator BAPTA-AM (10 μM, Cat# HY-100545, MCE).
Techniques: Fluorescence, Imaging, Membrane, Staining, Western Blot, Immunofluorescence, Expressing
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