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Files in this Data Supplement:
Fig S1. Regression plots showing dependence of HIF-1a protein amount on fish mass in crucian carp heart during normoxia and after 6, 24 and 48·h of hypoxia at 26°C (A–D), 18°C (E–H) and 8°C (I–L). Statistical significances were tested by linear regression analysis.
Fig S2. Electrophoretic shift analysis of HIF-1 DNA binding in nuclear extracts prepared from crucian carp gills. (A) The sequence specificity of HIF-1 DNA-binding. HIF-1 binds on HRE of human erythropoietin gene (wt) but not on mutated HRE of the same gene (m). (B) A supershift experiment shows that the presence of rainbow trout HIF-1a antibody (hif) in the reaction mixture inhibits formation of HIF-1-HRE complexes whereas the presence of actin-antibody (actin) or bovine serum albumin (bsa) in the reaction mixture does not affect HIF-1 DNA binding.
Fig S3. Northern blot analysis of total RNA isolated from the kidney of crucian carp. Each lane represents mRNAs of HIF-1a and b-actin and 18S ribosomal RNA from the kidney of one individual fish in normoxia at 8°C. 18S ribosomal RNA was used for normalization of the data.
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