Hypoxic HIF-1α protein accumulation and nuclear translocation in NB cells.
(A) Western blot analysis of HIF-1α was performed on whole-cell extracts from SH-SY5Y and Kelly cells cultured for 4 hours under normoxic (N), hypoxic (H), and anoxic (A) conditions. Antibody against α-tubulin was used to ensure equal loading. Gels are representative of at least 3 experiments. (B) To compare HIF-1α accumulation, HepG2 and SH-SY5Y cells were exposed to different O2 concentrations for 4 hours and whole-cell lysates (75 μg/lane) were subjected to Western blot analysis. (C) Indirect immunofluorescence analysis of HIF-1α in SH-SY5Y and HepG2 cells cultured for 4 hours under different O2 concentrations. Fluorescent intensities are shown in false colors, from blue (low fluorescence) to red (high fluorescence), as indicated by a colored scale bar; white scale bar represents 10 μm.
