SMC3 knockdown triggers genomic instability and p53-dependent apoptosis in human and zebrafish cells

Additional File 2:

Primers used for the semiquantitative analysis of the transcript level of selected zebrafish genes by RT-PCR. Oligonucleotide sequences matching those of the putative zebrafish genes were retrieved by querying the zebrafish dEST databases with tblastx and the protein sequence of the corresponding human genes. The retrieved zebrafish cDNA sequences were aligned and the longest open reading frame conceptually translated into the putative protein. The cDNA and the polypeptide sequence were in turn used to query the zebrafish genomic database to identify duplicated genes or repeated DNA sequences. Gene-specific PCR primers were designed to generate products of 500–800 bp.

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Additional File 1:

Effect of SMC3-siRNA on SMC3 and SMC1 protein level in human HCT116 cells. A) Western immunoblot assay of SMC3 and SMC1. Wild-type HCT116 cells were transfected with 50 ng/ml SMC3-siRNA and harvested 72 h later in lysis buffer. Fifty μg of cell lysate from either untransfected or transfected cells were separated on 7.5% SDS-PAGE and transferred by electroblotting to nitrocellulose filter. SMC3 and SMC1 were then detected using either goat anti-SMC3 or goat anti-SMC1 antibody followed by ECL reaction.

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