enzyme linked immunosorbent assay elisa  (Cusabio)

Journal: Scientific Reports

Article Title: IL-37 alleviates inflammatory effects and NLRP3 inflammasome activation in LPS-induced preterm birth

doi: 10.1038/s41598-025-34506-1

Figure Lengend Snippet: rhIL-37 delays preterm delivery by reducing LPS-induced inflammatory effects and NLRP3 inflammasome activation in mice. ( A ) Orbital blood was collected from the control mice ( n = 6), LPS-induced preterm mice (LPS; n = 6), and rhIL-37 + LPS-co-treated preterm mice (rhIL-37 + LPS; n = 6).Serum levels of IL-1β, IL-6 and TNF-α were detected via ELISA. ( B ) Foetal membrane and placental tissues were collected from mice ( n = 6 per group) immediately before labour,, and the mRNA expression levels of NLRP3, Caspase-1 and ASC were detected via qRT-PCR. ( C ) Foetal membrane and placental tissues were collected from the mice in each group ( n = 3 per group), and the protein levels of NLRP3, Caspase-1, Pro-caspase-1 and ASC were analyzed by western blotting. ( D ) Western blot Results were normalised to β-actin. Bar graphs show statistical quantification of the western blot data. All data met the assumption of homogeneity of variance. Data are presented as mean ± s.e.m. Results are representative of at least three independent experiments. * p < 0.05; ** p < 0.01;*** p < 0.001;**** p < 0.0001 (one-way ANOVA).

Article Snippet: Supernatants collected from HTR-8/SVneo cells were assayed for IL-1β, IL-6, and TNF-α levels using a human ELISA kit (LiankeBio, Hangzhou, China).

Techniques: Activation Assay, Control, Enzyme-linked Immunosorbent Assay, Membrane, Expressing, Quantitative RT-PCR, Western Blot

Journal: Scientific Reports

Article Title: IL-37 alleviates inflammatory effects and NLRP3 inflammasome activation in LPS-induced preterm birth

doi: 10.1038/s41598-025-34506-1

Figure Lengend Snippet: IL-37 knockdown enhances LPS-induced inflammatory and NLRP3 inflammasome activation in HTR-8/Svneo cells. ( A ) Cell supernatants were collected from control, si_NC- or si_IL37-treated cells following LPS exposure, and IL-37 expression level were detected by ELISA, qRT-PCR and Western blotting. ( B ) IL-1β, IL-6 and TNF-α levels in the cell supernatants were detected by ELISA. ( C ) The mRNA expression levels of NLRP3, Caspase-1 and ASC after cell treatment with LPS ( n = 5) were measured by qRT-PCR analysis. ( D ) NLRP3,pro-caspase-1, caspase-1 and ASC protein levels were detected by Western blotting, and the results were normalised to β-actin. All data exhibited equal variance.The data are shown as the mean ± s.e.m. The results are representative of at least three independent experiments. * p < 0.05; ** p < 0.01;*** p < 0.001;**** p < 0.0001 (One-way ANOVA).

Article Snippet: Supernatants collected from HTR-8/SVneo cells were assayed for IL-1β, IL-6, and TNF-α levels using a human ELISA kit (LiankeBio, Hangzhou, China).

Techniques: Knockdown, Activation Assay, Control, Expressing, Enzyme-linked Immunosorbent Assay, Quantitative RT-PCR, Western Blot

Journal: Scientific Reports

Article Title: IL-37 alleviates inflammatory effects and NLRP3 inflammasome activation in LPS-induced preterm birth

doi: 10.1038/s41598-025-34506-1

Figure Lengend Snippet: Nigericin reversed the anti-inflammatory effects and effects of rhIL-37 and its inhibition of NF-κB p65 activation in LPS-induced PTB.t ( A ) Cell supernatants were collected from HTR-8/Svneo cells co-treated with rhIL-37 and LPS, then either left untreated or treated with nigericin.The mRNA expression of NLRP3, caspase-1, and ASC was detected. ( B ) Cells were collected, and the protein expression of NLRP3, caspase-1, pro-caspase-1, and ASC was detected by western blotting. Results were normalised to β-actin and statistically analyzed.( C ) HTR-8/Svneo cell supernatants were collected after co-treatment with rhIL-37 and LPS (with or without nigericin), and the expression levels of IL-1β, IL-6, and TNF-α were detected by ELISA. ( D ) HTR-8/Svneo cells co-treated with rhIL-37 and LPS (with or without nigericin) were collected. The mRNA expression of p65 was detected by qRT-PCR, and protein expression of p65 and p-p65 was detected by western blotting. Densitometric quantification of p-p65 relative to p65 was performed. Statistical analysis of the western blot results was performed.All data exhibited equal variance.The data are shown as the means ± s.e.m. The results are representative of at least three independent experiments..

Article Snippet: Supernatants collected from HTR-8/SVneo cells were assayed for IL-1β, IL-6, and TNF-α levels using a human ELISA kit (LiankeBio, Hangzhou, China).

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

Journal: Aging Cell

Article Title: Periodic Therapeutic Phlebotomy Mitigates Systemic Aging Phenotypes by Promoting Bone Marrow Function

doi: 10.1111/acel.70400

Figure Lengend Snippet: PTP promotes the transformation of hematopoiesis to a youthful state by improving the hematopoietic microenvironment in the bone marrow. (A) Representative SA‐β‐gal staining of rat bone marrow (scale bars, 200 μm) (B) Quantification of SA‐β‐gal + cells per area, n = 3/group. (C) TGF‐β1 (pg/mL) and TPO (pg/mL) in rat bone marrow, n = 3/group. (D–N) The proportion of lin − cell subsets (HSC, LT‐HSC, ST‐HSC, MPP1, MPP5, MPP2, MPP3, CMP, MPP4, CLP, Pre GM, GMP, Pre MegE, MEP, MKP) in mice bone marrow cells, n = 3–4/group. Statistical analysis: One‐way ANOVA and Bonferroni post‐test. (O) Multiple cytokine profiling quantification of SASP proteins in mice bone marrow, log2‐transformed fold change in mean fluorescence intensity (MFI) compared to the average of D‐gal group, n = 6/group. (P) Representative examples and (Q) quantification of SEC (CD31 low Sca‐1 low ) and AEC (CD31 hi Sca‐1 hi ) in ECs from rat bone marrow and endosteum was detected by flow cytometry, n = 3/group. (R) The third generation of rat bone marrow‐derived BMSCs were cultured for 8 days. Cell viability was measured by MTT assay, n = 5/group. (S–T) Western blot of P16, P21, P27, and P63 expression in rat bone marrow‐derived BMSCs, n = 4/group. (U) Cell cycle analysis of rat bone marrow‐derived BMSCs by flow cytometry, n = 3/group. (V) Quantification of cell cycle phase distribution, n = 3/group. (W) Apoptosis of the third generation of rat bone marrow‐derived BMSCs using Annexin V‐FITC/PI KIT combined with flow cytometry, n = 3/group. Statistical analysis: One‐way ANOVA, Bonferroni post‐test for (D–N) and Tukey post‐test for the others, Data are mean ± SD; error bars denote 95% CI; * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: The concentration of Klotho (Elabscience, E‐EL‐R2580c), Taurine (Absin, abs580222), TGF‐β1 (Multi Sciences, EK981), TPO (Fine Test, ER0202) was measured in accordance with the protocols outlined in the respective reagent kit manuals.

Techniques: Transformation Assay, Staining, Fluorescence, Flow Cytometry, Derivative Assay, Cell Culture, MTT Assay, Western Blot, Expressing, Cell Cycle Assay