Fig. 5: Nociceptive sensory neuron-derived NGF mediates MSCs excessive fibrotic commitment.
A Prrx1Tomato mice experimental procedure and phenotypic diagram. B, C Double IF of αSMA (green) and Tomato (red) of peritendinous tissue from Prrx1Tomato mice intrathecal injected with AAV-GFP-siNGF or NC at 14 and 28 days after modeling (n = 6). Scale bars: 50 μm. D Transwell_co-culture strategy and immunofluorescent staining of Tomato (red) cells and α-SMA (green) cells from four groups in vitro. Scale bars: 20 μm. E, F Relative mRNA expression and protein expressions of Col1 and Col3 in cell experiment groups intrathecal injected with AAV-GFP-siNGF or NC (n = 3). G Double IF of CD31 (green) and Tomato (red) in Prrx1Tomato mice and NesTomato mice at 14 days after modeling (n = 6). Scale bars: low power image is 100 μm, and the high power image is 40 μm. H Double IF of αSMA (green) and Tomato (red) in Prrx1Tomato mice, αSMA (green) and Engrailed-1 (red) in WT mice at 14 days after modeling (n = 6). Scale bars: 50 μm. I Double IF of Engrailed-1 (green) and Tomato (red) in Prrx1Tomato mice at 14 days after modeling (n = 6). Scale bars: 50 μm. Yellow dashed line shows space between tendon and surrounding tissues. Black or white dashed line shows space occupied by fibrosis tissues. T represents tendon; AD represents fibrosis tissue; M represents muscle(mean ± SD; C/E–H: unpaired t-test; *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001; A(mouse, injector)/D(mouse) created in Adobe. Author, Yanhao Li. (2025). Contact the author for publication if needed; A(tendon, fibrosis tissue, MSCs and myofibroblasts)/D(MSCs and neurons) created in BioRender. Author, li, y. (2025)) https://BioRender.com/0hlz1hf).
