human nt 3  (Alomone Labs)

Journal: bioRxiv

Article Title: Fibroblasts neurotrophin signaling sustains pathological vascular maturation in rheumatoid arthritis

doi: 10.64898/2026.03.12.711120

Figure Lengend Snippet: (A-C) Bar plots showing differential regulation of pericyte markers (RGS5, ABCC9), VSMC markers (CNN1, MYH11), and pan-mural cell markers (ACTA2, SMTN, TAGLN) in fibroblasts treated with (A) NGF, (B) BDNF, or (C) NT3 compared with untreated fibroblasts. Bar heights represent log₂ fold change, and bar color indicates the corresponding P values. (D) Schematic of the collagen gel contraction assay in which synovial fibroblasts were embedded in collagen I gels and treatment (E) Representative images of collagen I gel contraction following treatment with NGF, BDNF, or NT3 (F) Quantification of collagen I gel contraction expressed as percent contraction. Values represent mean ± standard deviation (SD). Individual data points indicate biological replicates. Statistical analysis was performed using one-way ANOVA, with P values shown. (G) Schematic illustrating the synovial organoid/explant experimental design. (H) Representative H&E image of synovial explant culture after 3 days of culture. (I) Whole-mount immunoluorescence images showing preservation of endothelial cells after 3 days in culture, marked by CD31 (magenta), and vascular smooth muscle cells marked by MYH11, surrounding CD31 vascular structures. (J) Whole-mount staining showing expression of neurotrophin receptors NGFR, NTRK1, NTRK2, and NTRK3 (green) around vascularature, together with CD31 (magenta) and DAPI (blue). (K) Synovial organoids embedded in Matrigel and treated with NGF, BDNF, or NT3, followed by fixation and staining for a-smooth muscle actin (aSMA) (brown) to visualize mural cells. All 20X images were cropped at similar scale. Scale bar, 150 µm.

Article Snippet: For neurotrophin (NT) stimulation, recombinant NGF (256-GF, R&D Systems), BDNF (11166-BD, R&D Systems), and NT3 (267-N3-005, R&D Systems) were reconstituted in DMSO or PBS and diluted in media.

Techniques: Collagen Gel Contraction Assay, Standard Deviation, Preserving, Staining, Expressing

Journal: bioRxiv

Article Title: Fibroblasts neurotrophin signaling sustains pathological vascular maturation in rheumatoid arthritis

doi: 10.64898/2026.03.12.711120

Figure Lengend Snippet: (A) RNAscope images showing transcript expression of pericyte marker RGS5 (yellow) and VSMC marker MYH11 (red) in fibroblasts treated with different neurotrophins, overlaid with DAPI (blue). Scale bar, 150 µm. All images were acquired at 20x magnification and cropped to the same scale.(B) Quantification of total numbers of RGS5+, MYH11+, and RGS5+/MYH11+ double-positive cells following treatment with the indicated neurotrophins.(C) mRNA expression of the pericyte marker RGS5 in fibroblasts transfected with siRNAs targeting NGFR or NTRK1, in the presence or absence of NGF stimulation.(D) mRNA expression of the VSMC marker MYH11 in fibroblasts transfected with siRNAs targeting NTRK2 or NTRK3, in the presence or absence of BDNF or NT3 stimulation, respectively.(E-F) mRNA expression of the mural cell marker CNN1 (calponin) in fibroblasts stimulated with NGF (E) in the presence or absence of a TRKA inhibitor, or stimulated with BDNF (F) in the presence or absence of an NTRK2 inhibitor.

Article Snippet: For neurotrophin (NT) stimulation, recombinant NGF (256-GF, R&D Systems), BDNF (11166-BD, R&D Systems), and NT3 (267-N3-005, R&D Systems) were reconstituted in DMSO or PBS and diluted in media.

Techniques: RNAscope, Expressing, Marker, Transfection

Journal: bioRxiv

Article Title: Fibroblasts neurotrophin signaling sustains pathological vascular maturation in rheumatoid arthritis

doi: 10.64898/2026.03.12.711120

Figure Lengend Snippet: (A) Quantiication of a–smooth muscle actin (aSMA) staining in synovial organoids treated with neurotrophins (NGF, BDNF, NT3).(B) Cell viability assay of entrectinib and larotrectinib across a range of concentrations (0-100 uM). Values are presented as mean, with P values indicated (two-tailed student’s t-test). (C) Representative aSMA staining images of synovial organoids treated with neurotrophin receptor agonists. (D) Quantiication of aSMA staining following treatment with neurotrophin receptor agonists (LM22B-10 and 7,8dihydroxylavone).(E) Representative aSMA staining images of synovial organoids treated with neurotrophin receptor antagonists. (F) Quantiication of aSMA staining following treatment with neurotrophin receptor antagonists (DAPT, GNF5837, GW441756, and ANA-12). Values are presented as mean ± standard deviation (SD). Individual data points represent biological replicates, with P values indicated on the graphs. Statistical analysis was performed using one-way ANOVA for comparisons among multiple groups. Post hoc pairwise comparisons were conducted using t tests with Bonferroni correction to control multiple comparisons.

Article Snippet: For neurotrophin (NT) stimulation, recombinant NGF (256-GF, R&D Systems), BDNF (11166-BD, R&D Systems), and NT3 (267-N3-005, R&D Systems) were reconstituted in DMSO or PBS and diluted in media.

Techniques: Staining, Viability Assay, Two Tailed Test, Standard Deviation, Control