Skip to main content
Figure 1 | BMC Cancer

Figure 1

From: Aberrant over-expression of a forkhead family member, FOXO1A, in a brain tumor cell line

Figure 1

a) FOXO1A Northern analysis. mRNA isolated from the PER-452, PER-453, and PER-547 CNS-PNET cell lines was screened with a 32P labelled FOXO1A RT-PCR product spanning FOXO1A exons 1 and 2 including part of the forkhead domain (see Fig. 2b). A prominent band of ~5.9 kb, and a weaker band of ~5.4 kb were clearly visible in the PER-453 lane and are indicated by arrows on the upper right. Faint bands of similar sizes were visible in the PER-452 lane on the original autoradiograph and no signals in this size range were observed in the PER-547 lane. Molecular weight markers (nt) are included on the left. b) FOXO1A immunoblot. Total cell lysates from the PER-452, PER-453, and PER-480 CNS-PNET cell lines were screened with an antibody to FOXO1A. c) A ~78 kDa band, equivalent to the known size of wild type FOXO1A, is clearly visible in the PER-453 lane. A weak signal at 78 kDa was also detected in the PER-452 lane, and no signal was detected in the PER-480 lane. Equal proportions (~7%) of either the nuclear or cytoplasmic protein fractions isolated from the PER-453 cell line were also assessed with the FOXO1A antibody. FOXO1A protein was detected in both fractions with a greater proportion detectable in the nucleus. As fractionation controls the blot was re-probed with monoclonal antibodies to the predominantly cytoplasmic protein ACTB, and nuclear protein histone H1. Molecular weight markers (kDa) are included on the left. d) FOXO1A gene copy number analysis. Genomic DNA from the PER-453 cell line and two normal individuals was digested with EcoRI and analysed by Southern analysis using a FOXO1A specific cDNA probe. FOXO1A genomic fragments of 3320 bp and 613 bp are indicated by arrows in the top panel. To control for loading the blot was stripped and reprobed with a 32P labelled PCR product mapping to chromosome 14q that was expected to be present at diploid levels in the three specimens (bottom panel).

Back to article page