qPCR and western blot data confirm absence of Cx55.5 and Cx52.6.
(A) Immunoreactivity patterns of Cx55.5, Cx52.6, and Cx52.9 antibodies in membrane samples of wild-type and C54X mutant zebrafish retinas. On Western blots derived from 8–10% SDS-PA gradient gels on which membrane samples (40 µg of each) of wild-type and mutant zebrafish retinas were separated, the Cx55.5 and Cx52.6 detected a double band with a molecular weight in the expected range (black asterisk) in the wild-type, but not the mutant retina (black asterisk). In contrast, the expression of Cx52.9 protein appeared to be unaffected in the mutant (white asterisk) and a protein with the appropriate molecular weight of Cx52.9 was detected in both membrane samples, wild-type and mutant. (B) qPCR experiments revealed a significant down regulation of Cx55.5 and Cx52.6 mRNA in mutant fish (p<0.001). Steady state Cx52.9 mRNA levels were not affected.