Skip to main content
. 2004 Oct 12;32(18):5499–5511. doi: 10.1093/nar/gkh880

Figure 1.

Figure 1

Figure 1

Identification of a novel spliced variant of HIF-1α cDNA. (A) A comparison of the nucleotide sequences of the human EST clone (AW410666) and human HIF-1α cDNA. Dashed lines represent nucleotides deleted in the EST clone. Exons containing the HIF-1α and the EST clone are illustrated schematically in the lower panel. (B) Total RNA was extracted from Hep3B, HEK293, HeLa and SNU601 cells. HIF-1α cDNAs were amplified and identified using a highly sensitive RT–PCR method. The illustration in the right panel shows the nucleotide sequences of the exon 9/10 and exon 9/11 junctions. (C) The relative amounts of the mRNAs were calculated using the equation (c.p.m. of 250 bp-fragment/c.p.m. of 537 bp fragment) × (222/99). The correction factor, 222/99, represents the ratio of G and C numbers in the 537 bp fragment versus that of the 250 bp fragment. The results are representative of three separate experiments. (D) Filter membranes that contained 2 μg of human poly(A) mRNAs in each lane were purchased from Ambion Inc. (Austin, TX). Northern blot analysis was performed using 32P-labeled specific probes for HIF-1α and its variant. β-actin mRNA was analyzed as a loading control.