Fig 2: Tenascin C expression during palatal development in mice between embryonic day 13.5 (E13.5) and E15.5. (A) The mRNA expressions of tenascin-C (TNC), tenascin-X (TNX), and tenascin-W (TNW) in the anterior and posterior palatal shelves at E13.5, E14.5, and E15.5 (real-time polymerase chain reaction). The mRNA expression of TNC was greater than that of TNX or TNW and significantly higher in the posterior palate than in the anterior palate at E15.5. Data are presented as the mean ± standard deviation (n = 6). The mRNA expressions were normalized to that of glyceraldehyde 3-phosphate dehydrogenase (GAPDH). *P < 0.05. (B) Immunofluorescence staining of TNC protein in the middle, mid-posterior and posterior regions of the palatal shelves at E13.5, E14.0, E14.5, and E15.5. Stronger TnC expression was observed in the posterior palatal shelves than in the middle or mid-posterior regions, particularly at the later developmental stages. T, tooth germ; Pt, pterygoid process. Bar = 100 µm.

Fig 3: Transforming growth factor-beta-3 (TGF-ß3) and sonic hedgehog (SHH) increase the expression of tenascin-C (TNC) in mouse embryonic palatal mesenchymal (MEPM) cells (embryonic day 13.5–14.0). (A,C,E) The mRNA expression of tenascin-C (TNC) measured using real-time polymerase chain reaction. (B,D,F) The amount of TNC protein secreted into the culture medium measured by enzyme-linked immunosorbent assay. TGF-ß3 increased the expression of TNC mRNA (A) and the secretion of TNC protein (B) by MEPM cells at 24 h and 48 h. (C,D) These effects of TGF-ß3 (Tb3) were inhibited by a Smad3 inhibitor (Tb3 + SIS) and a p38 inhibitor (Tb3 + SB). SHH also enhanced the expression of TNC mRNA (E) and the secretion of TNC protein (F) by MEPM cells. Data are expressed as the mean ± standard deviation (n = 4) and were normalized to the 0 h or control values. *P < 0.05.

Fig 4: CAQK and tenascins in EAE lesions. (A–D) Confocal images identify FAM-CAQK peptide (A,E,I, green) at DAPI+ lesion clusters (B,F,J, blue) in the lumbar spinal cord of mice with EAE. The peptide does not associate with damaged myelin stained for MBP (C, magenta; merge in D) and largely complements staining for tenascin-R (G, magenta; merge in H). Some overlap is detected between FAM-CAQK and fibrous tenascin-C deposition (K, magenta) as indicated by the white signal in the merged image (L, arrowheads). Note that the majority of the lesion site covered by the peptide is negative for either tenascin (H,L). Scale bars: 20 µm.

Fig 5: Marginal CAQK peptide association with tenascins in the cuprizone model. Confocal images from the demyelinating corpus callosum of mice on a 6-week cuprizone diet show FAM-CAQK peptide (A,C, green) in complementary distribution with tenascin-R (B, magenta); merged images in (C). FAM-CAQK peptide (D) shows marginal association (white puncta pointed out by arrows in F) with tenascin-C (E magenta). DAPI-staining (blue) indicates cell nuclei in (C,F). D, dorsal; V, ventral; L, lateral; M, medial. Scale bar = 50 µm.