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Fig. 1
Inhibition of Cx43-GJIC recapitulates sofb123 phenotypes. (A) Schematic of Cx43 showing the location of Gap27 interaction with the second extracellular loop. N, amino terminus; C, carboxy terminus; EL1/EL2, extracellular loop1/2; CL, cytoplasmic loop. (B) Representative longitudinal cross-sections showing localization of 5FAM-Gap27 (green) following its injection. DAPI (blue) stains cell nuclei. m, mesenchyme; e, epidermis. UN, uninjected. (C) Regenerate length is reduced in Gap27-injected fins compared with Scrm-Gap27-injected fins. All fins were amputated at the 50% level. The amputation plane is indicated (black line). Black arrow indicates the distance from the amputation plane to the distal tip of the third fin ray. Injections were performed at 3 dpa and measurements were taken at 7 dpa. UN, uninjected. Bar graph (F) shows a significant reduction in the percentage similarity between Gap27 and Scrm-Gap27 (one-way ANOVA with Tukey's and Dunnett's multiple comparison as post-hoc tests; *P<0.05). n=21; three independent replicates. (D) Calcein-stained fin rays show reduced segment length in Gap27-injected fins compared with Scrm-Gap27-injected fins. Arrows and arrowheads point to the first two joints following the amputation plane (white line). The inset shows a higher magnification of the segments. Bar graph (G) reveals a significant reduction in the percentage similarity between Gap27 and Scrm-Gap27 (one-way ANOVA with Tukey's and Dunnett's multiple comparison as post-hoc tests; *P<0.05). n=19; three independent replicates. (E) H3P+ cells are reduced in number in Gap27-injected fins compared with Scrm-Gap27-injected fins. Arrowheads identify one H3P+ cell per image. Bar graph (H) reveals a significant reduction in the percentage similarity between Gap27 and Scrm-Gap27. Student's t-test, two tailed and unpaired, *P<0.05 (n=23, three independent replicates). Data are mean±s.e.m.