Journal: bioRxiv

Article Title: Endocrine-Adapted Pituitary Macrophages Regulate Gonadotropin Secretion through CXCL5-CXCR2-MAPK Signaling

doi: 10.64898/2026.08.12.744557

Figure Lengend Snippet: ( A ) LH secretion following 12- or 30-hour pre-stimulation with 100 ng/mL CXCL5 and subsequent 30-minute stimulation with 10 nM GnRH is presented as MFI. ( B - C ) Corresponding GnRH receptor (GnRHR) and CXCR2 surface expressions were measured by flow cytometry and expressed as MFI. ( D ) LβT2 cells were pre-treated with anti-CXCR2 antibody (1:250 dilution) for 30 minutes prior to 12-hour BMDM-cultured media treatment, with the antibody maintained at a final dilution of 1:500. LβT2 cells were co-cultured with BMDM-conditioned media following the density to volume ratio of 2 gonadotropes per 1 macrophage. LH concentration in culture supernatants was measured by Luminex. ( E ) Phospho-protein array analysis using a mouse MAPK signaling pathway kit. LβT2 cells were treated with 100 ng/mL CXCL5 for 30 minutes without prior serum starvation of cells.

Article Snippet: Antibody-printed membranes from the JAK/STAT and MAPK Phospho Antibody Array kits (RayBiotech, Cat. No. AAM-JAKSTAT-1-4, Cat. No. AAH-MAPK-1-4) were incubated with normalized cell lysates overnight at 4°C, followed by sequential incubation with biotinylated detection antibodies and HRP-conjugated streptavidin as recommended by the manufacturer.

Techniques: Flow Cytometry, Cell Culture, Concentration Assay, Luminex, Protein Array

Journal: Annals of Medicine

Article Title: Integrated multi-omics profiling of amniotic fluid identifies predictive biomarkers for fetal growth restriction trajectories

doi: 10.1080/07853890.2026.2718562

Figure Lengend Snippet: External validation of diagnostic and prognostic biomarkers. (A,B) Absolute concentrations of (A) PDGFA and (B) phospho-STAT5A (Tyr694) in the FGR and control groups. (C) ROC curve analysis of PDGFA, phospho-STAT5A (Tyr694), and their combined diagnostic model for FGR identification. (D–F) Absolute concentrations of (D) IREB2, (E) HLA-C, and (F) PLXNB2 across the transient and persistent FGR groups. (G) ROC curve analysis of IREB2, HLA-C, PLXNB2, and their combined model for prognostic stratification.

Article Snippet: The concentrations of functional PDGF-AA homodimer (Catalog No. ELH-PDGFAA, RayBiotech, USA) and the activation levels of phosphorylated STAT5A at the Tyr694 site [phospho-STAT5A (Tyr694)] (Catalog No. PEL-STAT5A-Y694-T, RayBiotech, USA) were quantified via multi-step sandwich ELISA according to the manufacturer’s specific protocols.

Techniques: Biomarker Discovery, Diagnostic Assay, Control

Journal: Journal of Neuroscience Research

Article Title: miR ‐19a‐3p and miR ‐19b‐3p Promote Microglia Activation Associated With Neuroinflammation

doi: 10.1002/jnr.70137

Figure Lengend Snippet: miR‐19a and miR‐19b increased phosphorylation of NFκB p65 in microglia. Microglia were transfected with miR‐19a mimic or miR‐19b mimic. Microglia were stimulated with LPS and transfected with miR‐19a mimic, miR‐19b mimic, or negative control miR mimic. After 24 h, the microglia were lysed and whole protein extracts were analyzed by ELISA for P65 (serine 536). Significant difference (*) was determined by one way anova and Bonferroni's multiple comparison test (* p < 0.001) based on unstimulated microglia. These are representative graphs from one experiment of three independent repeated experiments.

Article Snippet: Following incubation for 24 h, the amount of phosphorylated NFκB p65 Ser536 protein in the microglia cell lysate was measured using commercial mouse‐specific ELISA kits (RayBiotech) according to the manufacturer's instructions.

Techniques: Phospho-proteomics, Transfection, Negative Control, Enzyme-linked Immunosorbent Assay, Comparison