e el m0048  (Elabscience Biotechnology)

Journal: iScience

Article Title: PBX3 regulates mast cell parthanatos via TOP2A mediated DNA damage in allergic rhinitis

doi: 10.1016/j.isci.2026.115426

Figure Lengend Snippet: Regulatory role of TOP2A in mast cell apoptosis, parthanatos pathway, and inflammatory response In vitro experimental groups: CON, model, Model+OE-NC, Model+OE-TOP2A, Model+si-NC, Model+si-TOP2A. (A and B) RT-qPCR and western blot analyses were conducted to assess the mRNA and protein expression levels of TOP2A and PARP-1 in mast cells. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001 vs. the CON or Model+OE-NC or Model+si-NC group. (C) TUNEL staining was performed to evaluate and visualize cell apoptosis. ∗ p < 0.05, ∗∗ p < 0.01 vs. the CON or Model+OE-NC or Model+si-NC group; scale bars, 50 μm. (D) Cell proliferation activity in each group was measured using the CCK-8 assay. ∗∗ p < 0.01 vs. the CON or Model+OE-NC or Model+si-NC group. (E and F) MMP was assessed using the JC-1 assay to evaluate mitochondrial function. ∗∗ p < 0.01, ∗∗∗ p < 0.001 vs. the CON or Model+OE-NC or Model+si-NC group. (G) Western blotting was used to examine the protein levels of mitochondrial AIF (Mito-AIF), cytoplasmic AIF (Cyto-AIF), and nuclear AIF (Nucleo-AIF), along with visual analysis. ∗∗ p < 0.01, ∗∗∗ p < 0.001 vs. the CON or Model+OE-NC or Model+si-NC group. (H and I) ELISA was employed to determine the concentrations of β-hexosaminidase, Histamine, IL-4, IL-5, and IFN-γ. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001 vs. the CON or Model+OE-NC or Model+si-NC group. Data are represented as mean ± SEM ( n = 3 per group) from independent biological replicates. One-way ANOVA with Tukey’s post hoc correction for (A–D) and (F–I).

Article Snippet: IFN-γ ELISA Kit , Elabscience , Cat# E-EL-M0048.

Techniques: In Vitro, Quantitative RT-PCR, Western Blot, Expressing, TUNEL Assay, Staining, Activity Assay, CCK-8 Assay, Enzyme-linked Immunosorbent Assay

Journal: iScience

Article Title: PBX3 regulates mast cell parthanatos via TOP2A mediated DNA damage in allergic rhinitis

doi: 10.1016/j.isci.2026.115426

Figure Lengend Snippet: Effects of TOP2A on DNA damage, parthanatos regulation, and degranulation in mast cells Experimental groups: Model+OE-NC, Model+OE-TOP2A, Model+si-NC, Model+si-TOP2A. (A) Western blotting was employed to determine the protein expression levels of γ-H2AX. ∗∗ p < 0.01, ∗∗∗ p < 0.001 vs. the Model+OE-NC or Model+si-NC group. (B and C) IF staining was performed to assess the expression of the DNA damage marker γ-H2AX in mast cells. ∗∗ p < 0.01, ∗∗∗ p < 0.001 vs. the Model+OE-NC or Model+si-NC group; scale bar = 50 μm. (D) Cellular senescence was evaluated using an SA-β-gal staining kit. ∗ p < 0.05, ∗∗ p < 0.01 vs. the Model+OE-NC or Model+si-NC group; scale bars, 50 μm. Experimental groups: Model+OE-NC, Model+OE-TOP2A, Model+OE-TOP2A + DOX. (E and F) RT-qPCR and western blot analyses were conducted to detect the gene and protein expression levels of TOP2A and PARP-1 under different treatment conditions. ∗∗∗ p < 0.001 vs. the Model+OE-NC or Model+OE-TOP2A group; ns, not significant ( p ≥ 0.05). (G) Western blotting was employed to determine the protein expression levels of γ-H2AX. ∗∗∗ p < 0.001 vs. the Model+OE-NC or Model+OE-TOP2A group. (H) IF staining was used to evaluate γ-H2AX expression as a marker of DNA damage. ∗ p < 0.05, ∗∗∗ p < 0.001 vs. the Model+OE-NC or Model+OE-TOP2A group; scale bars, 50 μm. (I) SA-β-gal staining was used to assess cellular senescence. ∗ p < 0.05, ∗∗ p < 0.01 vs. the Model+OE-NC or Model+OE-TOP2A group; scale bars, 50 μm. (J) Western blot analysis was used to determine the protein expression levels of Mito-AIF, Cyto-AIF, and Nucleo-AIF. ∗∗ p < 0.01, ∗∗∗ p < 0.001 vs. the Model+OE-NC or Model+OE-TOP2A group. (K and L) ELISA assays were carried out to measure the levels of β-hexosaminidase, Histamine, IL-4, IL-5, and IFN-γ. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001 vs. the Model+OE-NC or Model+OE-TOP2A group. Data are represented as mean ± SEM ( n = 3 per group) from independent biological replicates. one-way ANOVA with Tukey’s post hoc correction for (A and C–L).

Article Snippet: IFN-γ ELISA Kit , Elabscience , Cat# E-EL-M0048.

Techniques: Western Blot, Expressing, Staining, Marker, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay

Journal: iScience

Article Title: PBX3 regulates mast cell parthanatos via TOP2A mediated DNA damage in allergic rhinitis

doi: 10.1016/j.isci.2026.115426

Figure Lengend Snippet: Effects of PBX3 silencing on mast cell function and DNA damage response Experimental groups: Model+si-NC, Model+ si-PBX3. (A) RT-qPCR and western blot analyses were conducted to evaluate the mRNA expression levels of PBX3 and TOP2A. ∗∗∗ p < 0.001 vs. the Model+si-NC group. (B–D) Western blotting and IF staining were used to assess the expression of the DNA damage marker γ-H2AX. ∗ p < 0.05, ∗∗∗ p < 0.001 vs. the Model+si-NC group; scale bars, 50 μm. (E) SA-β-gal staining was performed to detect cellular senescence. ∗∗ p < 0.01 vs. the Model+si-NC group; scale bars, 50 μm. (F–H) RT-qPCR, western blotting, and IF staining were employed to examine PARP-1 expression in mast cells. ∗∗∗ p < 0.001 vs. the Model+si-NC group; scale bars, 50 μm. (I) Western blot analysis was used to detect the protein levels of Mito-AIF, Cyto-AIF, and Nucleo-AIF. ∗∗∗ p < 0.001 vs. the Model+si-NC group. (J and K) ELISA assays were conducted to quantify the levels of β-hexosaminidase, histamine, IL-4, IL-5, and IFN-γ in mast cells. ∗∗ p < 0.01, ∗∗∗ p < 0.001 vs. the Model+si-NC group. Data are represented as mean ± SEM ( n = 3 per group) from independent biological replicates. Two-tailed unpaired Student’s t tests for (A, B, D–G, and I–K).

Article Snippet: IFN-γ ELISA Kit , Elabscience , Cat# E-EL-M0048.

Techniques: Cell Function Assay, Quantitative RT-PCR, Western Blot, Expressing, Staining, Marker, Enzyme-linked Immunosorbent Assay, Two Tailed Test

Journal: iScience

Article Title: PBX3 regulates mast cell parthanatos via TOP2A mediated DNA damage in allergic rhinitis

doi: 10.1016/j.isci.2026.115426

Figure Lengend Snippet: PBX3 drives DNA damage through TOP2A to regulate mast cell parthanatos, senescence, and degranulation Experimental groups: Model+si-NC, Model+si-PBX3, Model+si-PBX3+OE-NC, Model+si-PBX3+OE-TOP2A. (A) Western blot analysis was performed to determine protein expression levels of PBX3, TOP2A, PARP-1, and γ-H2AX. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001 vs. the Model+si-NC or Model+si-PBX3+OE-NC group; ns, not significant ( p ≥ 0.05). (B) IF staining was conducted to detect the DNA damage marker γ-H2AX. ∗∗∗ p < 0.001 vs. the Model+si-NC or Model+si-PBX3+OE-NC group; scale bars, 50 μm. (C) Cellular senescence was assessed using an SA-β-gal staining kit. ∗∗∗ p < 0.001 vs. the Model+si-NC or Model+si-PBX3+OE-NC group; scale bars, 50 μm. (D) ELISA was employed to measure the levels of degranulation markers β-hexosaminidase and histamine. ∗∗ p < 0.01, ∗∗∗ p < 0.001 vs. the Model+si-NC or Model+si-PBX3+OE-NC group. (E) Cytokine levels of IL-4, IL-5, and IFN-γ in cell supernatants were quantified by ELISA. ∗ p < 0.05, ∗∗∗ p < 0.001 vs. the Model+si-NC or Model+si-PBX3+OE-NC group. Data are represented as mean ± SEM ( n = 3 per group) from independent biological replicates. One-way ANOVA with Tukey’s post hoc correction for (A–E).

Article Snippet: IFN-γ ELISA Kit , Elabscience , Cat# E-EL-M0048.

Techniques: Western Blot, Expressing, Staining, Marker, Enzyme-linked Immunosorbent Assay

Journal: iScience

Article Title: PBX3 regulates mast cell parthanatos via TOP2A mediated DNA damage in allergic rhinitis

doi: 10.1016/j.isci.2026.115426

Figure Lengend Snippet: Regulatory role of TOP2A in mast cell apoptosis, parthanatos pathway, and inflammatory response In vitro experimental groups: CON, model, Model+OE-NC, Model+OE-TOP2A, Model+si-NC, Model+si-TOP2A. (A and B) RT-qPCR and western blot analyses were conducted to assess the mRNA and protein expression levels of TOP2A and PARP-1 in mast cells. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001 vs. the CON or Model+OE-NC or Model+si-NC group. (C) TUNEL staining was performed to evaluate and visualize cell apoptosis. ∗ p < 0.05, ∗∗ p < 0.01 vs. the CON or Model+OE-NC or Model+si-NC group; scale bars, 50 μm. (D) Cell proliferation activity in each group was measured using the CCK-8 assay. ∗∗ p < 0.01 vs. the CON or Model+OE-NC or Model+si-NC group. (E and F) MMP was assessed using the JC-1 assay to evaluate mitochondrial function. ∗∗ p < 0.01, ∗∗∗ p < 0.001 vs. the CON or Model+OE-NC or Model+si-NC group. (G) Western blotting was used to examine the protein levels of mitochondrial AIF (Mito-AIF), cytoplasmic AIF (Cyto-AIF), and nuclear AIF (Nucleo-AIF), along with visual analysis. ∗∗ p < 0.01, ∗∗∗ p < 0.001 vs. the CON or Model+OE-NC or Model+si-NC group. (H and I) ELISA was employed to determine the concentrations of β-hexosaminidase, Histamine, IL-4, IL-5, and IFN-γ. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001 vs. the CON or Model+OE-NC or Model+si-NC group. Data are represented as mean ± SEM ( n = 3 per group) from independent biological replicates. One-way ANOVA with Tukey’s post hoc correction for (A–D) and (F–I).

Article Snippet: Histamine ELISA Kit , Elabscience , Cat# E-EL-M0048.

Techniques: In Vitro, Quantitative RT-PCR, Western Blot, Expressing, TUNEL Assay, Staining, Activity Assay, CCK-8 Assay, Enzyme-linked Immunosorbent Assay

Journal: iScience

Article Title: PBX3 regulates mast cell parthanatos via TOP2A mediated DNA damage in allergic rhinitis

doi: 10.1016/j.isci.2026.115426

Figure Lengend Snippet: Effects of TOP2A on DNA damage, parthanatos regulation, and degranulation in mast cells Experimental groups: Model+OE-NC, Model+OE-TOP2A, Model+si-NC, Model+si-TOP2A. (A) Western blotting was employed to determine the protein expression levels of γ-H2AX. ∗∗ p < 0.01, ∗∗∗ p < 0.001 vs. the Model+OE-NC or Model+si-NC group. (B and C) IF staining was performed to assess the expression of the DNA damage marker γ-H2AX in mast cells. ∗∗ p < 0.01, ∗∗∗ p < 0.001 vs. the Model+OE-NC or Model+si-NC group; scale bar = 50 μm. (D) Cellular senescence was evaluated using an SA-β-gal staining kit. ∗ p < 0.05, ∗∗ p < 0.01 vs. the Model+OE-NC or Model+si-NC group; scale bars, 50 μm. Experimental groups: Model+OE-NC, Model+OE-TOP2A, Model+OE-TOP2A + DOX. (E and F) RT-qPCR and western blot analyses were conducted to detect the gene and protein expression levels of TOP2A and PARP-1 under different treatment conditions. ∗∗∗ p < 0.001 vs. the Model+OE-NC or Model+OE-TOP2A group; ns, not significant ( p ≥ 0.05). (G) Western blotting was employed to determine the protein expression levels of γ-H2AX. ∗∗∗ p < 0.001 vs. the Model+OE-NC or Model+OE-TOP2A group. (H) IF staining was used to evaluate γ-H2AX expression as a marker of DNA damage. ∗ p < 0.05, ∗∗∗ p < 0.001 vs. the Model+OE-NC or Model+OE-TOP2A group; scale bars, 50 μm. (I) SA-β-gal staining was used to assess cellular senescence. ∗ p < 0.05, ∗∗ p < 0.01 vs. the Model+OE-NC or Model+OE-TOP2A group; scale bars, 50 μm. (J) Western blot analysis was used to determine the protein expression levels of Mito-AIF, Cyto-AIF, and Nucleo-AIF. ∗∗ p < 0.01, ∗∗∗ p < 0.001 vs. the Model+OE-NC or Model+OE-TOP2A group. (K and L) ELISA assays were carried out to measure the levels of β-hexosaminidase, Histamine, IL-4, IL-5, and IFN-γ. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001 vs. the Model+OE-NC or Model+OE-TOP2A group. Data are represented as mean ± SEM ( n = 3 per group) from independent biological replicates. one-way ANOVA with Tukey’s post hoc correction for (A and C–L).

Article Snippet: Histamine ELISA Kit , Elabscience , Cat# E-EL-M0048.

Techniques: Western Blot, Expressing, Staining, Marker, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay

Journal: iScience

Article Title: PBX3 regulates mast cell parthanatos via TOP2A mediated DNA damage in allergic rhinitis

doi: 10.1016/j.isci.2026.115426

Figure Lengend Snippet: Effects of PBX3 silencing on mast cell function and DNA damage response Experimental groups: Model+si-NC, Model+ si-PBX3. (A) RT-qPCR and western blot analyses were conducted to evaluate the mRNA expression levels of PBX3 and TOP2A. ∗∗∗ p < 0.001 vs. the Model+si-NC group. (B–D) Western blotting and IF staining were used to assess the expression of the DNA damage marker γ-H2AX. ∗ p < 0.05, ∗∗∗ p < 0.001 vs. the Model+si-NC group; scale bars, 50 μm. (E) SA-β-gal staining was performed to detect cellular senescence. ∗∗ p < 0.01 vs. the Model+si-NC group; scale bars, 50 μm. (F–H) RT-qPCR, western blotting, and IF staining were employed to examine PARP-1 expression in mast cells. ∗∗∗ p < 0.001 vs. the Model+si-NC group; scale bars, 50 μm. (I) Western blot analysis was used to detect the protein levels of Mito-AIF, Cyto-AIF, and Nucleo-AIF. ∗∗∗ p < 0.001 vs. the Model+si-NC group. (J and K) ELISA assays were conducted to quantify the levels of β-hexosaminidase, histamine, IL-4, IL-5, and IFN-γ in mast cells. ∗∗ p < 0.01, ∗∗∗ p < 0.001 vs. the Model+si-NC group. Data are represented as mean ± SEM ( n = 3 per group) from independent biological replicates. Two-tailed unpaired Student’s t tests for (A, B, D–G, and I–K).

Article Snippet: Histamine ELISA Kit , Elabscience , Cat# E-EL-M0048.

Techniques: Cell Function Assay, Quantitative RT-PCR, Western Blot, Expressing, Staining, Marker, Enzyme-linked Immunosorbent Assay, Two Tailed Test

Journal: iScience

Article Title: PBX3 regulates mast cell parthanatos via TOP2A mediated DNA damage in allergic rhinitis

doi: 10.1016/j.isci.2026.115426

Figure Lengend Snippet: PBX3 drives DNA damage through TOP2A to regulate mast cell parthanatos, senescence, and degranulation Experimental groups: Model+si-NC, Model+si-PBX3, Model+si-PBX3+OE-NC, Model+si-PBX3+OE-TOP2A. (A) Western blot analysis was performed to determine protein expression levels of PBX3, TOP2A, PARP-1, and γ-H2AX. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001 vs. the Model+si-NC or Model+si-PBX3+OE-NC group; ns, not significant ( p ≥ 0.05). (B) IF staining was conducted to detect the DNA damage marker γ-H2AX. ∗∗∗ p < 0.001 vs. the Model+si-NC or Model+si-PBX3+OE-NC group; scale bars, 50 μm. (C) Cellular senescence was assessed using an SA-β-gal staining kit. ∗∗∗ p < 0.001 vs. the Model+si-NC or Model+si-PBX3+OE-NC group; scale bars, 50 μm. (D) ELISA was employed to measure the levels of degranulation markers β-hexosaminidase and histamine. ∗∗ p < 0.01, ∗∗∗ p < 0.001 vs. the Model+si-NC or Model+si-PBX3+OE-NC group. (E) Cytokine levels of IL-4, IL-5, and IFN-γ in cell supernatants were quantified by ELISA. ∗ p < 0.05, ∗∗∗ p < 0.001 vs. the Model+si-NC or Model+si-PBX3+OE-NC group. Data are represented as mean ± SEM ( n = 3 per group) from independent biological replicates. One-way ANOVA with Tukey’s post hoc correction for (A–E).

Article Snippet: Histamine ELISA Kit , Elabscience , Cat# E-EL-M0048.

Techniques: Western Blot, Expressing, Staining, Marker, Enzyme-linked Immunosorbent Assay

Journal: Virulence

Article Title: Wzi modulates immune evasion of hypervirulent Klebsiella pneumoniae in a CPS-dependent and independent manner

doi: 10.1080/21505594.2026.2656049

Figure Lengend Snippet: The influence of wzi on IFN-γ related cytokines secretion. (a-c) Mice ( n = 8–10) were intranasally infected with 1 × 10 5 CFU wt (red) and Δ wzi (blue) mutant strains. Lungs were homogenated for measuring IL-12 (a), IFNα (b) and IFN-γ (c) secretions at 5 and 10 hpi using ELISA, and results were normalized to the protein concentration. (d-f) mice ( n = 8–10) were intranasally infected with 1 × 10 5 CFU wt (red), Δ wcaJ (yellow) and Δ wcaJ / wzi (purple) mutants. Lungs were homogenated for measuring IL-12 (d), IFNα (e) and IFN-γ (f) secretions at 10 hpi using ELISA, and results were normalized to the protein concentration. Each data point represents an individual animal. (g) BMDMs were pretreated with or without IFN-γ (20 ng/mL) for 12 h, then infected with K. pneumoniae for 1 h (MOI = 50:1). Wells were washed three times with PBS, and incubated with RPMI1640 + 10% FBS medium containing gentamicin (100 mg/L). At the indicated time, intracellular bacteria were quantified by 5% saponin, serial dilution and viable count on lb agar plates. Each assay was performed in triplicate. (a-f) Kruskal – Wallis tests followed by Dunn’s post-hoc tests with Holm’s correction were applied, (g) two-tailed Mann–Whitney U tests were used. *, p < 0.05; **, p < 0.01; ***, p < 0.001.

Article Snippet: The ELISA kits (Elabscience) included mouse IFN-γ ELISA kit (E-EL-M0048); mouse IL-12 ELISA kit (E-EL-M3062); mouse IFN-α ELISA kit (E-EL-M3054); mouse CXCL1 ELISA kit (E-EL-M0018).

Techniques: Infection, Mutagenesis, Enzyme-linked Immunosorbent Assay, Protein Concentration, Incubation, Bacteria, Serial Dilution, Two Tailed Test, MANN-WHITNEY