HIF-1α protein and HIF target gene expression in ρ0 cell lines.
(A,C,D) Immunoblots showing levels of HIF-1α in cell extracts from wild-type and mtDNA-less cell lines. Cells were cultured in parallel for 4 hours in normoxia (21% O2, N) or hypoxia (0.1% O2, H). (A) Human osteosarcoma 143B and the derived mtDNA-less 206ρ0 cell line. (B) Human osteosarcoma 143B and the derived 206ρ0 cell line. (C) Human lung cancer cell line A549 and the derived mtDNA-less B2ρ0 cell line. (D) Further extracts of normoxic A549 and B2ρ0 cells. βTubulin acts as an internal control. (B,E) Expression of Glut-1 mRNA. Cells were cultured in parallel for 16 hours in normoxia (21% O2, N) or hypoxia (0.1% O2, H). RNAse protection assays of total RNA was performed for Glut-1 and U6 small nuclear RNA as an internal control. (E) A549 and the derived B2ρ0 cell line.
