lactate dehydrogenase ldh activity assay kit  (Elabscience Biotechnology)

Journal: bioRxiv

Article Title: Lipid A counteracts doxorubicin-induced systemic dysfunction by boosting mitochondrial activity

doi: 10.64898/2026.04.16.719094

Figure Lengend Snippet: (A, B) LVEF (A) and LVFS (B) 4 weeks after adjuvant administration. (C, D) Body weight (C) and heart weight (D) 4 weeks after adjuvant administration (n = 4-6 mice per group). (E, F) Cytotoxicity in NRCMs 20 hours after adjuvant treatment. Cell viability and toxicity were measured by LDH release and MTT assay (n = 3). All data are shown as mean ± SEM; Data were analyzed using one-way ANOVA followed by Tukey’s comparison test.

Article Snippet: Twenty hours after drug treatment, cytotoxicity and cell viability were evaluated using the Cytotoxicity LDH Assay Kit-WST (Dojindo Molecular Technologies, Inc., Kumamoto, Japan) and the Cell Counting Kit-8 (Dojindo Molecular Technologies, Inc., Kumamoto, Japan), respectively, in accordance with the manufacturers’ manuals.

Techniques: Adjuvant, MTT Assay, Comparison

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 Letters

Article Title: Lung squamous cell carcinoma downregulates haptoglobin expression to inhibit M2 macrophage ferroptosis via the hemoglobin-dependent CD163/HO-1 pathway

doi: 10.3892/ol.2026.15558

Figure Lengend Snippet: Overexpression of HP in LUSC cells does not induce ferroptosis in LUSC cells. (A and B) NCI-H226 and NCI-H520 cells were infected with LV- HP or LV-Ctrl. After 72 h of infection, cells and culture supernatants were collected for analysis of HP (A) mRNA and (B) protein levels by RT-PCR and ELISA, respectively. (C) NCI-H226 and NCI-H520 cells infected with LV- HP or LV-Ctrl were cultured with or without Hb (30 µg/ml) for 24 h. Culture supernatants were collected and LDH release was measured. (D) Cells were analyzed via flow cytometry to determine the percentage of 7-AAD-positive cells in NCI-H226 and NCI-H520 cells. (E and F) The mean fluorescence intensity of (E) FerroOrange (Fe 2+ ) and (F) C11-BODIPY (lipid peroxide) was measured using flow cytometry. Data are presented as the mean ± SD from five independent experiments and were analyzed using an unpaired t-test or a two-way ANOVA followed by a Tukey's post-hoc multiple comparison analysis. ****P<0.0001. ns, not significant; HP, haptoglobin; Hb, hemoglobin; LDH, lactate dehydrogenase; 7-AAD, 7-aminoactinomycin D.

Article Snippet: An LDH cytotoxicity assay kit (cat. no. HY-K1090; MedChemExpress) was used to assess LDH levels in the supernatant of LUSC cell lines (NCI-H226 and NCI-H520 cells) and M2 macrophages, according to the manufacturer's protocol.

Techniques: Over Expression, Infection, Reverse Transcription Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay, Cell Culture, Flow Cytometry, Fluorescence, Comparison

Journal: Oncology Letters

Article Title: Lung squamous cell carcinoma downregulates haptoglobin expression to inhibit M2 macrophage ferroptosis via the hemoglobin-dependent CD163/HO-1 pathway

doi: 10.3892/ol.2026.15558

Figure Lengend Snippet: CM from LUSC cells overexpressing HP leads to ferroptosis in M2 macrophages. (A) THP-1 cells were differentiated into macrophages by treating them with 150 nM PMA for 24 h, followed by culture in fresh RPMI-1640 for 24 h. The macrophages were polarized into M2 macrophages by incubating with IL-4 (20 ng/ml) and IL-13 (20 ng/ml) for 24 h. Flow cytometry was then performed to identify CD163 + CD206 + M2 macrophages. (B) M2 macrophages were cultured with the CM from NCI-H226 and NCI-H520 cells infected with LV- HP or LV-Ctrl in the presence or absence of Hb for 24 h. LDH release was analyzed by ELISA. Treated M2 macrophages were treated as in (B) and flow cytometry was used to detect the proportion of (C) 7-AAD positive cells, as well as the mean fluorescence intensity of (D) FerroOrange and (E) C11-BODIPY. (F) Treated M2 macrophages were observed using transmission electron microscopy. Scale bar, 5 µm. Data are presented as the mean ± SD from five independent experiments and were analyzed using an unpaired t-test or a two-way ANOVA followed by a Tukey's post-hoc multiple comparison analysis. ****P<0.0001. ns, not significant; CM, conditioned media; LUSC, lung squamous cell carcinoma; LDH, lactate dehydrogenase; 7-AAD, 7-aminoactinomycin D; HP, haptoglobin.

Article Snippet: An LDH cytotoxicity assay kit (cat. no. HY-K1090; MedChemExpress) was used to assess LDH levels in the supernatant of LUSC cell lines (NCI-H226 and NCI-H520 cells) and M2 macrophages, according to the manufacturer's protocol.

Techniques: Flow Cytometry, Cell Culture, Infection, Enzyme-linked Immunosorbent Assay, Fluorescence, Transmission Assay, Electron Microscopy, Comparison