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( A ) Experimental design of anti-LOX treatment in KPC mice. ( B ) LOX activity after early anti-LOX and late anti-LOX treatment. ( C ) Representative histology images, obtained by Trichrome stain, picrosirius red (including polarized images and collagen fiber individualization analysis), and scanning electron microscope (SEM). Scale bar, 1 and 50 μm as indicated. ( D ) Quantification of Trichrome (collagen content). ( E ) Quantification of collagen fiber width, length, and straightness. ( F ) Frequency distribution of collagen fiber diameter (selected from 5 fields of SEM images). ( G ) Young’s modulus of fresh tumor samples, measured by atomic force microscopy. ( H ) Representative images of PGP9.5 + nerve fibers in tumor tissues (left panel; scale bar, 50 μm). Quantification of PGP9.5 + nerve area (right panel). ( I ) 3D images of a pancreas from KPC mice labeled with anti-PGP9.5 and anti-CK19. ( J ) Axon projections in PDAC and PanIN lesions used for quantification of nerve densities. Scale bar, 30 μm. In A - G , n = 8 for control, n = 7 for early anti-LOX, and n = 7 for late anti-LOX. * P < 0.05, ** P < 0.01 and *** P < 0.001. Values as mean ± SD and compared by the one-way ANOVA multiple comparisons with Tukey’s method among groups.

Journal: bioRxiv

Article Title: Mechanical cues of extracellular matrix determine tumor innervation

doi: 10.1101/2024.03.25.586535

Figure Lengend Snippet: ( A ) Experimental design of anti-LOX treatment in KPC mice. ( B ) LOX activity after early anti-LOX and late anti-LOX treatment. ( C ) Representative histology images, obtained by Trichrome stain, picrosirius red (including polarized images and collagen fiber individualization analysis), and scanning electron microscope (SEM). Scale bar, 1 and 50 μm as indicated. ( D ) Quantification of Trichrome (collagen content). ( E ) Quantification of collagen fiber width, length, and straightness. ( F ) Frequency distribution of collagen fiber diameter (selected from 5 fields of SEM images). ( G ) Young’s modulus of fresh tumor samples, measured by atomic force microscopy. ( H ) Representative images of PGP9.5 + nerve fibers in tumor tissues (left panel; scale bar, 50 μm). Quantification of PGP9.5 + nerve area (right panel). ( I ) 3D images of a pancreas from KPC mice labeled with anti-PGP9.5 and anti-CK19. ( J ) Axon projections in PDAC and PanIN lesions used for quantification of nerve densities. Scale bar, 30 μm. In A - G , n = 8 for control, n = 7 for early anti-LOX, and n = 7 for late anti-LOX. * P < 0.05, ** P < 0.01 and *** P < 0.001. Values as mean ± SD and compared by the one-way ANOVA multiple comparisons with Tukey’s method among groups.

Article Snippet: Primary antibodies were S100β (66616-1-Ig, Proteintech, 1:200), NF-L (ab223343, Abcam, 1:100), Collagen I (#72026, Cell Signaling Technology, 1:200), LOX (17958-1-AP, Proteintech, 1:200), TGM2 (15100-1-AP, Proteintech, 1:200), α-SMA (A5228, Sigma-Aldrich, 1:400), PGP9.5 (14730-1-AP, Proteintech, 1:200), PGP9.5 (66230-1-Ig, Proteintech, 1:500; for co-IF analysis), CK19 (1:200, Proteintech, 10712-1-AP), and AMY2A (15845-1-AP, Proteintech, 1:100).

Techniques: Activity Assay, Staining, Microscopy, Labeling