Journal: Bone & Joint Research

Article Title: Piezo1 drives fibroblast activation in epidural fibrotic remodelling via the ET-1/HIF-1α pathway

doi: 10.1302/2046-3758.158.BJR-2025-0662.R1

Figure Lengend Snippet: Ca² + and endothelin-1 (ET-1) contribute to Piezo1-dependent activation of hypoxia-inducible factor 1-alpha (HIF-1α). a) and b) Gene set enrichment analysis (GSEA) of epidural scar tissue from mice after laminectomy. c) Ca² + influx measured by Fluo-4 AM in the blank, type I rat tail collagen-encapsulated, and collagen-encapsulated with GsMTX4 groups. d) Western blot analysis of NIH/3T3 fibroblasts treated with type I rat tail collagen and the calcium influx blocker Nimodipine (n = 3). e) Western blot analysis of NIH/3T3 fibroblasts treated with Yoda1 and Nimodipine (n = 3). f) Quantitative analysis of relative Edn1 messenger RNA (mRNA) expression in blank and collagen-encapsulated NIH/3T3 fibroblasts. g) and h) Western blot analysis of NIH/3T3 fibroblasts treated with type I rat tail collagen and the ET-1 inhibitor Bosentan (n = 3). *p < 0.05, **p < 0.01, ***p < 0.001, independent-samples t -test (two groups) or analysis of variance (multiple groups).

Article Snippet: Alternatively, the following drugs were used to stimulate NIH/3T3 cells: Yoda1 (25 μM; HY-P1410), a chemical activator of Piezo1; BAY 87-2243 (1 μM; HY-15836), an inhibitor of hypoxia-inducible factor 1-alpha (HIF-1α); SB-431542 (10 μM; HY-10431), an inhibitor of TGF-β1; SIS 3 (10 μM; HY-13013), an inhibitor of Smad3; GsMTX4 (2.5 μM; HY-P1410); Nimodipine (60 μM; HY-B0265), a calcium channel blocker; and Bosentan (1 μM; HY-A0013, all MedChemExpress), an endothelin-1 (ET-1) receptor antagonist.

Techniques: Activation Assay, Western Blot, Expressing

Journal: Brain

Article Title: Post-mortem evidence of pathogenic angiogenesis and abnormal vascular function in early Alzheimer’s disease

doi: 10.1093/brain/awaf394

Figure Lengend Snippet: Elevated VEGF-A and EDN1 associated with cerebral hypoperfusion in early-stage Alzheimer's disease. Concentrations of ( A ) VEGF-A and ( B ) EDN1, measured by sandwich ELISA, in Braak tangle stage groups BS0–II, BSIII–IV and BSV–VI, across brain regions. Each data-point represents the average of a duplicate measurement in a single brain region. Z -scores were calculated to combine data from two independent cohorts. Mean ± standard error of the mean are shown. Correlation analysis between ( C ) VEGF-A and ( D ) EDN1 with the MAG:PLP1 ratio in the different brain regions. Solid linear regression lines are used to indicate brain regions with significant correlations, dashed lines indicate brain regions with non-significant relationships. Trigone is not shown as it did not fit within the range of the graph. * P < 0.05, ** P < 0.01, *** P < 0.001 indicate statistical strength of correlations within respective brain regions.

Article Snippet: The EDN1 level was measured using a commercially available ELISA kit (QET00B, R&D systems), as reported previously., Brain tissue homogenates were diluted 1:50 in assay diluent and 100 μl was added to the plate in duplicate.

Techniques: Sandwich ELISA

Journal: Brain

Article Title: Post-mortem evidence of pathogenic angiogenesis and abnormal vascular function in early Alzheimer’s disease

doi: 10.1093/brain/awaf394

Figure Lengend Snippet: Mediators of angiogenesis are dysregulated in early-stage/intermediate pathology Alzheimer's disease. ( A and B ) Levels of endoglin (CD105), a marker of neoangiogenesis, measured first using the angiogenesis proteome profiler and then independently by commercial ELISA kit. Mean and standard error of the mean are shown. * P < 0.05, ** P < 0.01. ( C and D ) Levels of pro-angiogenic ( C ) and anti-angiogenic ( D ) mediators in early-stage/intermediate pathology [Braak tangle stage (BS) III–IV] (blue) and late-stage AD (BSV–VI) (red), relative to the levels in minimal pathology BS0–II cases.

Article Snippet: The EDN1 level was measured using a commercially available ELISA kit (QET00B, R&D systems), as reported previously., Brain tissue homogenates were diluted 1:50 in assay diluent and 100 μl was added to the plate in duplicate.

Techniques: Marker, Enzyme-linked Immunosorbent Assay