Journal: Chembiochem

Article Title: Functional Heterogeneity of the Autism Spectrum Disorder‐Associated Gut Microbial Ecosystem Revealed by Fecal Lipopolysaccharides and Bacterial Proteomic Profiling

doi: 10.1002/cbic.70557

Figure Lengend Snippet: Effects of fecal LPS on HMC3 microglia cell viability and activation. (A) Bright‐field image of HMC3 cells (40× magnification, scale bar: 20 µm). (B) HMC3 microglia cell viability after 24 h exposure to increasing concentrations (0.1, 1, 10, 100 ng/mL) of the pooled fecal LPS preparations obtained from the ASD and NASD cohorts. Cell viability was expressed as % relative to non‐stimulated (NS) cells and assessed by MTT assay (570 nm). NS cells were also considered as negative control. (C,D) Quantification of (C) IL‐8 and (D) IL‐6 levels following LPSs treatment, measured by DuoSet‐ELISA kit. Three independent experiments ( n = 3) were performed, with each experimental condition tested in triplicate wells within each experiment. Statistical significance was assessed by ordinary one‐way ANOVA followed by Tukey's multiple‐comparison test. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001 versus NS; # p < 0.05, ### p < 0.001, #### p < 0.0001, versus LPS.

Article Snippet: Cytokine levels (pg/mL) in HMC3 supernatants were measured using DuoSet ELISA kits (R&D Systems, Minneapolis, MN, USA) for human IL‐8 (#DY208) and IL‐6 detection (#DY206).

Techniques: Activation Assay, MTT Assay, Negative Control, Enzyme-linked Immunosorbent Assay, Comparison

Journal: Frontiers in Immunology

Article Title: The role of NEDD4-mediated ubiquitination of AQP5 in allergic rhinitis: implications for nasal mucosal barrier function

doi: 10.3389/fimmu.2026.1908261

Figure Lengend Snippet: Disruption of the NEDD4-AQP5 axis impairs barrier function and promotes cytokine release. (A) MUC5AC amounts (pg/mL) in cellular culture supernatants were assayed with ELISA across various treatment groups. (B) TER (Ω·cm²) of HNEpC monolayers was measured utilizing a Millicell ERS-2 system across the specified groups. (C, D) The amounts of IL-6 (pg/mL) and IL-8 (pg/mL) in cellular culture supernatants were determined via ELISA. Data are shown as mean ± SD from three independent experiments, with each sample measured in triplicate. *P < 0.05, **P < 0.01, ***P < 0.001 vs. Ctrl group; # P < 0.05, ## P < 0.01, ### P < 0.001 compared with the HDM group. Ctrl, untreated control; HDM, house dust mite-sensitized HNEpCs; HDM + siNEDD4, HDM-sensitized HNEpCs transfected with siNEDD4; HDM + ovAQP5, HDM-sensitized HNEpCs transfected with AQP5 overexpression plasmid.

Article Snippet: Cell culture supernatants were used to detect IL-6, IL-8, and MUC5AC levels in the supernatant by using ELISA kits for IL-6 (R&D Systems, D6050), IL-8 (R&D Systems, D8000C), and MUC5AC (R&D Systems, DY2197).

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

Journal: Frontiers in Immunology

Article Title: ANKRD22 contributes to NF-κB-associated inflammatory responses in alveolar type II epithelial cells during PM 2.5 -triggered airway inflammation

doi: 10.3389/fimmu.2026.1894350

Figure Lengend Snippet: ANKRD22 inhibition mitigated PM 2.5 -induced inflammation in A549 cells. (A, B) A549 cells were transfected with ANKRD22 siRNA then stimulated with PM 2.5 , mRNA expression (A) and protein levels (B) of ANKRD22 were determined by qRT-PCR and Western blot. (C–G) mRNA expression levels of IL6 (C) , IL8 (D) , IL1B (E) , CXCL1 (F) , and CXCL2 (G) in A549 cells were detected by qRT-PCR. (H, I) ELISA measurement of IL-6 (H) and IL-8 (I) protein levels in A549 cells culture supernatants. Multiple comparisons were performed using one-way ANOVA followed by Bonferroni’s post hoc test. Experimental data analysis results are expressed as mean ± SEM. Data are representative of three biological replicates with similar results (n = 3 per group) ; * P < 0.05, ** P < 0.01, *** P < 0.001. ns, non-significant.

Article Snippet: In vitro experiment, the culture supernatants from A549 cells were collected, and the protein level of IL-6 (RK00004, ABclonal, China) and IL-8 (RK00011, ABclonal, China) were measured by ELISA.

Techniques: Inhibition, Transfection, Expressing, Quantitative RT-PCR, Western Blot, Enzyme-linked Immunosorbent Assay, Analysis