Fig 1: Immunofluorescence staining of connexins (Cxs): Cx37 and Cx40 in wild type and yotari mouse liver at gestation days E13.5 and E15.5. Immunoexpression of Cx37, 4′,6-diamidino-2- phenylindole dihydrochloride (DAPI) staining and merged Cx37 and DAPI at E13.5 and E15.5 in wild type (a) and yotari (b). Immunoexpression of Cx40, DAPI staining and merged Cx40 and DAPI at E13.5 and E15.5 in wild type (c) and yotari (d). The scale bar is 8 µm, refers to all images.
Fig 2: The area percentages of connexins (Cxs): Cx26, Cx32, Cx37, Cx40, Cx43, Cx45, pannexin1 (Panx1) and apoptosis-inducing factor (AIF) in wild type and yotari livers at gestation days E13.5 and E15.5. Data are presented as the mean ± SD (vertical line). Significant differences were indicated by * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001. One-way ANOVA followed by Tukey’s multiple comparisons test.
Fig 3: Co-expression of different Cxs with markers of proximal tubules (DBA) and distal tubules (LTL) in developing, postnatal and CNF kidneys. (a–d) Human kidneys in the 10th developmental week. (e–h,i–l) Human postnatal kidneys (1,5 years). (m–p) Postnatal CNF kidneys (3,5 years). Cx37, Cx40 and Cx45 expression (arrows), expression of markers for proximal (DBA) and distal (LTL) tubules (arrows), proximal tubules (pt), distal tubules (dt), collecting tubules (ct), interstitium (i), glomeruli (g), blood vessels (bv). Co-expression of Cx37 with LTL is missing in 10th developmental week kidneys (a–d), while co-expression of Cx40/DBA (arrowhead) (e–h) and Cx43/LTL (arrowhead) (i–l) is seen in proximal and distal tubules of postnatal kidneys. In CNF, Cx37 co-expresses with DBA (arrowhead) in dilated proximal tubules. Double immunofluorescence staining of DAPI with Cx37/LTL, Cx40/DBA, Cx43/LTL and Cx37/DBA x40.
Fig 4: Expression of connexin protein in cumulus-oocyte complexes (COCs). (A) Western blot image of connexin 37 (CX37) and the corresponding GAPDH as the loading control. (B) Densitometric analysis of CX37 protein expression relative to GAPDH. (C) Western blot image of connexin 43 (CX43) and corresponding GAPDH. (D) Densitometric analysis of CX43 and relative GAPDH protein. Proteins were extracted from groups of 40 COCs after 22 h of maturation in one of the five treatment groups. Six biological replicates for CX37 and 3 biological replicates for CX43 were analyzed. Bars represent mean ± SEM.
Fig 5: Expression of connexin 37 (Cx37) in the rat spinal cord. (a–h) Dorsal horn; (i) central canal (CC). Thoracic segments of the spinal cord were stained for Cx37 (green; arrows) and glial fibrillary acidic protein (gfap) (red; arrows). Blue—4′,6-diamidino-2-phenylindole (DAPI). (a–c) Lower magnification (scale bar on c = 50 µm); (i) scale bar = 20 µm; (d–h) and detailed inset on (i) higher magnification (scale bar on f = 10 µm). Arrowheads represent co-localization of Cx37 and gfap (yellow); asterisks show neuronal nuclei; bv denotes blood vessels.
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