nerve growth factor ngf  (Alomone Labs)

Journal: bioRxiv

Article Title: Decoding the fibroblast/mast cell signaling pathway of acupuncture

doi: 10.1101/2025.11.20.688774

Figure Lengend Snippet: a. Schematic of stretched fibroblasts on mast cells through paracrine signaling. Substance P (SP) and 5-hydroxytryptamine (5-HT) released by P815 mast cells were measured after culturing in the supernatant of stretched fibroblasts (5%; 0, 0.5 or 2 Hz; 30 minutes) for 4 hours (n = 6). b. Schematic of mechanical force on P815 cells, co-cultured with fibroblasts. SP and 5-HT levels in the co-culture supernatant after stretching (5%; 0, 0.5 or 2 Hz; 30 minutes) (n = 6). c. Schematic of the Transwell device (left): P815 cells were seeded in the upper chamber, with the lower chamber containing complete medium (Ctrl), conditioned medium from non-stretched fibroblasts (No stretch), or conditioned medium from stretched fibroblasts (Stretch). Images of cell clustering were stained with crystal violet after 8, 12 or 16 hours of incubation (middle). Scale bar: 50 μm. Statistical chart of mast cells under chemotaxis (n = 6) (right). d. Schematic of cytokine levels in the supernatant of fibroblasts after stretching by ELISA (left). NGF/CXCL1/M-CSF/IL-33/SCF levels measured in cell-free supernatants of fibroblasts after stretching (5%; 2 Hz) for 30 minutes (n = 6). e. Immunofluorescence staining of mice skin primary fibroblasts, with and without Y-27632, pre- and post-stretch: phalloidin (red), nucleus (blue), IL-33 or SCF (green). f. Fluorescence intensity statistics of IL-33 (top) and SCF (down) in fibroblasts. For IL-33, Ctrl (n = 507), Stretch (n = 493), Y-27632 with Stretch (n = 452). For SCF, Ctrl (n = 255), Stretch (n = 359), Y-27632 with Stretch (n = 404). Scale bars are 50 μm.

Article Snippet: The level of 5-HT, SP, NGF, M-CSF, CXCL1, IL-33, and SCF were analyzed using ELISA kits (E-EL-0033c, E-EL-0067c, E-EL-M0815, E-MSEL-M0105, E-EL-M0018, E-EL-M2642c, and E-EL-M0636c, respectively) from Elabscience, following manufacturer’s instructions.

Techniques: Cell Culture, Co-Culture Assay, Staining, Incubation, Chemotaxis Assay, Enzyme-linked Immunosorbent Assay, Immunofluorescence, Fluorescence

Journal: Journal of Bone and Mineral Research

Article Title: Pain in fibrous dysplasia: identifying nociceptive mechanisms in a preclinical model

doi: 10.1093/jbmr/zjaf039

Figure Lengend Snippet: In vitro cAMP assay (A), NGF ELISA (B), and multiplex assay (C-K). FD cells and control cells were activated with doxycycline (10 μg/mL) or vehicle (phosphate buffered saline) for 24 hr with IBMX. Intracellular cAMP was assessed using cAMP Gs dynamic kit and protein factors were assessed using MSD U-PLEX assay. Two-way ANOVA with Tukey’s multiple comparison test. Graphs are individual values with bars representing median ± IQR. **** p < .0001. Abbreviations: cAMP, cyclic adenosine monophosphate; FD, fibrous dysplasia; IBMX, 3-isobutyl-1-methylxanthine; NGF, nerve growth factor.

Article Snippet: cAMP concentration assay (Cisbio GS Dynamic Kit, PerkinElmer), Meso Scale Discovery Multiplex assay (U-PLEX; MSD), and nerve growth factor (NGF) ELISA (NGF Rapid ELISA kit: mouse; Avantor) were conducted according to manufacturer directions.

Techniques: In Vitro, cAMP Assay, Enzyme-linked Immunosorbent Assay, Multiplex Assay, Control, Saline, Plex Assay, Comparison