qpcr  (Biotium)

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: Mitochondrial Membrane Potential Detection Kit (JC-1) , Beyotime , Cat# C2006.

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