Journal: Current Therapeutic Research, Clinical and Experimental

Article Title: The Berberine Derivative BBR684 Inhibits VDAC Oligomerization to Suppress Ferroptosis in Acute Kidney Injury

doi: 10.1016/j.curtheres.2026.100825

Figure Lengend Snippet: BBR684 attenuates Erastin-induced ferroptosis in HK-2 cells by suppressing lipid peroxidation and activating the antioxidant response. (A) Flow cytometric analysis of lipid peroxidation using the C11-BODIPY probe in HK-2 cells treated as indicated for 12 hours. Data are presented as mean fluorescence intensity (MFI). (B) Intracellular Fe²⁺ levels measured by a ferrous iron colorimetric assay. (C, D) Levels of malondialdehyde (MDA, C) and reduced glutathione (GSH, D) in cell lysates. (E, F) Representative immunofluorescence images (E) and quantification (F) of 4-hydroxynonenal (4-HNE) adducts (red) in HK-2 cells. Nuclei were counterstained with DAPI (blue). Scale bar, 50 µm. (G) Western blot analysis of key ferroptosis-related proteins: glutathione peroxidase 4 (GPX4), heme oxygenase-1 (HO-1), and nuclear factor erythroid 2–related factor 2 (NRF2). β-actin served as the loading control. Data are from 3 independent biological replicates (n = 3) and presented as mean ± SEM. Significance was determined by one-way ANOVA followed by Tukey's HSD post hoc test. * P < 0.05, ** P < 0.01, *** P < 0.001; n.s., not significant.

Article Snippet: Intracellular Fe2+ levels were quantified using a Ferrous Iron Colorimetric Assay Kit (Catalog #E-BC-K773-M, Elabscience, Wuhan, China) per the manufacturer's protocol.

Techniques: Fluorescence, Colorimetric Assay, Immunofluorescence, Western Blot, Control

Journal: Current Therapeutic Research, Clinical and Experimental

Article Title: The Berberine Derivative BBR684 Inhibits VDAC Oligomerization to Suppress Ferroptosis in Acute Kidney Injury

doi: 10.1016/j.curtheres.2026.100825

Figure Lengend Snippet: Overexpression of VDAC abolishes the protective effects of BBR684 against ferroptosis.(A) Western blot analysis validating the overexpression of VDAC1 in HEK293T cells. (B) Representative phase-contrast microscopy images showing the morphology of HEK293T cells with or without VDAC1 overexpression after the indicated treatments for 24 hours. (C) Quantification of cell death by flow cytometry (propidium iodide staining) in HEK293T cells treated with Erastin and BBR684. (D) Cells viability assessed by the CCK-8 assay in HEK293T cells treated with increasing concentrations of BBR684 for 24 hours. (n = 3, 3 biological replicates). (E, F) Levels of malondialdehyde (MDA, F) and reduced glutathione (GSH, E) in cell lysates. (n = 3, 3 biological replicates). (G) Intracellular Fe²⁺ levels measured by a colorimetric assay. (n = 3, 3 biological replicates).

Article Snippet: Intracellular Fe2+ levels were quantified using a Ferrous Iron Colorimetric Assay Kit (Catalog #E-BC-K773-M, Elabscience, Wuhan, China) per the manufacturer's protocol.

Techniques: Over Expression, Western Blot, Microscopy, Flow Cytometry, Staining, CCK-8 Assay, Colorimetric Assay