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qPCR and western blot data confirm absence of Cx55.5 and Cx52.6.

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posted on 19.07.2011, 01:37 by Lauw J. Klaassen, Ziyi Sun, Marvin N. Steijaert, Petra Bolte, Iris Fahrenfort, Trijntje Sjoerdsma, Jan Klooster, Yvonne Claassen, Colleen R. Shields, Huub M. M. Ten Eikelder, Ulrike Janssen-Bienhold, Georg Zoidl, Douglas G. McMahon, Maarten Kamermans

(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.

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