Fig 2: Immunofluorescence microscopy of LPC mice validation of shotgun proteomics analysis. (A) Representative images of LPC mice from the sedentary control, exercising animals, and secondary control for connexin-32 (CX32), and myelin (MBP). Scale bar denotes 100 µm. (B) Quantification of lesion area. (C) Quantification of percent of lesion that is MBP-positive. *p < 0.05. (D) Quantification of percent of lesion that is CX32-positive. **p < 0.01. A Mann–Whitney test was used for statistics. Each point is of a single animal, and the bar represents mean ± SD (n of 8 per group).

Fig 3: IF analysis for Cx26, Cx32, and Cx43 in NS-hLSGBs. (A–C) Representative confocal microscopy images of double IF with transmitted light contrast. Merged images show Cx positivity in red, ASMA positivity in green, and Cx-ASMA colocalization in yellow. These images have been modified by enhancing the saturation in order to better appreciate the yellow stain. White arrows indicate red immunoreaction at the plasma membrane of the duct cells and at the basal/basolateral side of the acinar cells. Black arrows point to the yellow spot due to the colocalization of Cx and ASMA. Inserts show the higher magnification of the squares. (D–F) Single IF images representing Cx immunoreactions (red) corresponding to (A–C) microscopic fields. A, acinus; D, duct. Scale bars, 20 µm; Inserts, 10 µm.

Fig 4: TEM immunogold analysis for Cx32 in NS-hLSGB. Representative images of immunoreactions (arrows) at the level of different cell compartments: rough endoplasmic reticulum and Golgi complex (A), mitochondria (B), nucleus (C), and apparent GJ in the lateral plasma membranes (D) of mucous cells. In (E) gold particles are visible on the apical luminal membrane of a duct cell and on a GJ between the lateral plasma membranes of two duct cells. D, desmosome; GC, Golgi complex; L, lumen; M, mitochondrium; MSG, mucous secretory granule; N, nucleus; RER, rough endoplasmic reticulum. Scale bars (A,C,E), 500 nm; (B) and (D), 200 nm.

Fig 5: IF analysis for Cx26, Cx32, and Cx43 in SS-hLSGBs. (A–C) Representative confocal microscopy images of double IF with transmitted light contrast. Merged images show Cx positivity in red, ASMA positivity in green, and Cx-ASMA colocalization in yellow. These images have been modified by enhancing the saturation in order to better appreciate the yellow stain. White arrows indicate red immunoreaction at the plasma membrane of the duct cells and at the basal/basolateral side of the acinar cells. Black arrows point to the yellow spot due to the colocalization of Cx and ASMA. Inserts show the higher magnification of the squares. (D–F) Single IF images representing Cx immunoreactions (red) corresponding to (A–C) microscopic fields. Asterisks (*) point to the inflammatory cells. A, acinus; D, duct. Scale bars, 20 µm.