atp content chemiluminescence assay kit  (Elabscience Biotechnology)

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

Journal: International Journal of Molecular Medicine

Article Title: Triptolide exerts antiviral effects and alleviates influenza A-induced pneumonia by inhibiting the overactivation of absent in melanoma 2 signaling in immune cells

doi: 10.3892/ijmm.2026.5829

Figure Lengend Snippet: Effects of TP on AIM2 signaling, LDH leakage, and pyroptosis in HBEpiCs and THP-1 cells. (A) RT-qPCR analysis revealed that TP treatment did not alter AIM2 mRNA expression in HBEpiCs but markedly reduced AIM2 mRNA levels in THP-1 cells. (B) TP did not affect the LDH leakage rate in HBEpiCs but markedly reduced LDH leakage in THP-1 cells. (C) Dual-fluorescence staining revealed that TP treatment decreased the formation of ASC-AIM2 complexes in THP-1 cells (scale bar, 10 μ m). (D) Western blot analysis indicated that TP treatment did not markedly affect the expression of these proteins in HBEpiCs. (E) TP treatment reduced the protein expression of AIM2, caspase-1, and GSDMD in THP-1 cells. (F) Flow cytometry revealed no difference in cell death between HBEpiCs treated with 20 nM TP and the control. (G) In THP-1 cells infected with H1N1 and treated with 20 nM TP, a significant reduction in cell death was identified compared with that in the H1N1 infection group. The data are presented as the mean ± standard deviation; * P<0.05, ** P<0.01, *** P<0.001 vs. control. TP, triptolide; AIM2, absent in melanoma 2; LDH, lactate dehydrogenase; HBEpiCs, human bronchial epithelial cells; RT-qPCR, reverse transcription-quantitative PCR; H1N1, influenza A; GSDMD, gasdermin D; Con, control.

Article Snippet: After treatment, 50 μ l of the supernatant from each well was transferred to a new plate, and an equal volume of LDH reaction mix from the Lactate Dehydrogenase (LDH) Activity Assay Kit (cat. no. E-BC-K046-M; Elabscience Bionovation Inc.) was added.

Techniques: Quantitative RT-PCR, Expressing, Fluorescence, Staining, Western Blot, Flow Cytometry, Control, Infection, Standard Deviation, Reverse Transcription, Real-time Polymerase Chain Reaction

Journal: International Journal of Molecular Medicine

Article Title: Triptolide exerts antiviral effects and alleviates influenza A-induced pneumonia by inhibiting the overactivation of absent in melanoma 2 signaling in immune cells

doi: 10.3892/ijmm.2026.5829

Figure Lengend Snippet: AIM2 overexpression reverses the immunosuppressive effects of TP in THP-1 cells. (A) Transfection with Ad-AIM2 upregulated the expression of AIM2 compared with that of Ad-NC in THP-1 cells. (B) AIM2 protein levels were elevated in THP-1 cells overexpressing AIM2 compared with control THP-1 cells. (C) In THP-1 cells, AIM2 overexpression increased the LDH leakage rate. (D) AIM2 overexpression reversed the TP-induced reduction in adhesion between THP-1 cells and HBEpiCs (scale bar, 10 μ m). (E) AIM2 overexpression enhanced the migration ability of THP-1 cells compared with that of H1N1-treated control cells and reversed the TP-induced decrease in THP-1 cell migration (scale bar, 50 μ m). (F) AIM2 overexpression reversed the TP-induced reduction in inflammatory cytokine levels. The data are presented as the mean ± standard deviation; * P<0.05, ** P<0.01, *** P<0.001 vs. Con; # P<0.05, ## P<0.01, ### P<0.001 vs. Ad-AIM2. AIM2, absent in melanoma 2; TP, triptolide; LDH, lactate dehydrogenase; HBEpiCs, human bronchial epithelial cells; H1N1, influenza A; GSDMD, gasdermin D; IL, interleukin; TNF-α, tumor necrosis factor-α; Con, control.

Article Snippet: After treatment, 50 μ l of the supernatant from each well was transferred to a new plate, and an equal volume of LDH reaction mix from the Lactate Dehydrogenase (LDH) Activity Assay Kit (cat. no. E-BC-K046-M; Elabscience Bionovation Inc.) was added.

Techniques: Over Expression, Transfection, Expressing, Control, Migration, Standard Deviation

Journal: Oncology Reports

Article Title: PINK1-mediated mitophagy enhances breast cancer proliferation through metabolic reprogramming

doi: 10.3892/or.2026.9117

Figure Lengend Snippet: Analysis of glycolysis-associated metabolite level changes in MCF-7 and MDA-MB-231 breast cancer cells. Levels of (A) PGK2, (B) pyruvate and (C) acetyl-CoA in MCF-7 cells, and (D) PGK2, (E) pyruvate and (F) acetyl-CoA in MDA-MB-231 cells transfected with vector control, PINK1-OE or PINK1-OE + siPGK2. Quantitative analysis was performed using ELISA in cells under the three indicated conditions. Statistical analysis was performed using a one-way ANOVA followed by Tukey's post-hoc test. Data are represented as the mean ± SEM from at least three independent experiments. ***P<0.001. PGK2, phosphoglycerate kinase 2; PINK1, PTEN-induced kinase 1; OE, overexpression; si, small interfering.

Article Snippet: Acetyl-CoA content was assessed using the Acetyl-CoA ELISA Kit (Wuhan Elabscience Biotechnology Co., Ltd.; cat. no. E-BC-F046-48T-ELS).

Techniques: Transfection, Plasmid Preparation, Control, Enzyme-linked Immunosorbent Assay, Over Expression