Knockdown of DJ-1 in zebrafish embryos. A. Abundant DJ-1 expression in human brain lysate from a control case, as detected by Western Blot (WB) with KAM-SA100 and anti-DJ-1-N antibodies, but not when anti-DJ-1-N was pre-absorbed with the synthetic DJ-1 peptide. Specificity of anti-DJ-1-N antibody was confirmed by detecting DJ-1 in wild type mouse brain lysate but not in the DJ-1 knockout mouse brain lysate. A minor cross-reacting band at ∼70 kDa was detected in extracts of both wild type and DJ-1 knockout mouse brain, but not in human brain extracts. B. Zebrafish embryos at different developmental stages and brains of adult zebrafish were lysed for Western blotting with antibody E2.19. High levels of DJ-1 protein were found in embryos at all developmental stages and in adult brains. C. Extracts taken from individual embryos at 24 hpf were run on WB with antibody E2.19 (bottom panel). The same blot was re-probed with antibody against α-tubulin (top panel). DJ-1 protein levels were dramatically reduced in DJ-1 KD embryos compared to control embryos.
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