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. 2024 Apr 25;14:9497. doi: 10.1038/s41598-024-59713-0

Figure 7.

Figure 7

Immunocytochemical features of gingival connective tissues in the gingiva of RNS variants. (A) In normal gingiva, POSTN is localized in a loose extracellular network around blood vessels (arrowheads; bv) whereas in both RNS-1 (B) and RNS-2 (C) mutants POSTN extracellular expression is localized along thick and disorganized fibers in the connective tissue (ct). (D) In normal gingiva, Collagen 5 alpha-1 (green; Col5A1) is secreted and deposited along collagen 1 fibers and Vimentin (red; Vim) is expressed by endothelial cells. (E,F) Increased expression of Col5A1 is detected in the connective tissue (ct) of RNS-1 (E) and RNS-2 (F) mutants.Vim is normally localized in endothelial cells and abnormally in fibroblasts (F). (G) Alpha-SMA (α-SMA) is a specific marker of contractile endothelial cells of bv. (H,I) In RNS mutants, α-SMA is also expressed by fibroblasts in ct. Note the presence of inflammatory infiltrates (i) in RNS-1 gingiva. (J) In normal gingiva, Fibroblast-specific protein 1 (green; FSP1) is expressed by gingival fibroblasts (green arrow). Osteonectin (red; SPARC) is expressed in discrete spots by fibroblasts (red arrowhead) and endothelial cells. (K,L) FSP-1 is normally expressed in ct. SPARC expression and SPARC deposition along fibers are dramatically increased in RNS mutant connective tissues. (M) Fibronectin (green; FN) is specifically expressed by fibroblasts. Phospho-SMAD3 (red; pSMAD3) is mainly localized in nuclei of vascular cells (arrowhead). (N,O) In RNS-derived gingiva FN expression is dramatically decreased. pSMAD3 expression is dramatically increased in both vascular cells (arrowhead) and fibroblasts (arrows) nuclei. Scale bars: (AC), (GL) = 100 μm; (DF), (MO) = 30 μm.