Journal: The Journal of Neuroscience

Article Title: Lack of Hypoxia-Inducible Factor-1α Impairs Midbrain Neural Precursor Cells Involving Vascular Endothelial Growth Factor Signaling

doi: 10.1523/JNEUROSCI.2482-06.2007

Figure Lengend Snippet: Degeneration of the SN of HIF-1α CKO mice. A, Immunoblots demonstrating TH and ALDH1A1 protein expression, neuronal β-tubulin III (TUJ1) expression, apoptotic effectors such as Bcl-2, pro-caspase-3, and activated caspase-3, and HIF-1α downstream target genes such as VEGF, all parallel assessed in the SN of HIF-1α CKO and cre/wt mice. B, SN of HIF-1α CKO mice viewed by confocal microscopy showing colocalization of activated caspase-3 (CM1; red) in TH-immunoreactive cells (green). C, Stereological investigation revealing the number of dopaminergic (TH-positive) neurons in the SN and the ventral tegmental area of HIF-1α CKO in relation to cre/wt mice is presented. Scale bar, 40 μm. Data are expressed as mean ± SEM values from five independent experiments. *p < 0.05 when compared with cre/wt NPCs.

Article Snippet: Antibodies used to probe blots were as follows: mouse monoclonal anti-proliferating cell nuclear antigen (PCNA) (Santa Cruz Biotechnology, Santa Cruz, CA); mouse monoclonal anti-Bcl-2 (Santa Cruz Biotechnology), mouse monoclonal anti-pro-caspase-3 (BD Transduction Laboratories, San Jose, CA); rabbit polyclonal anti-activated caspase-3 (CM1; BD PharMingen, San Diego, CA); mouse monoclonal anti-VEGF (Abcam, Cambridge, UK); chicken polyclonal anti-aldehyde dehydrogenase (ALDH1A1) (kindly provided by N. E. Sladek, Minneapolis, MN); mouse monoclonal anti-actin (MP Biomedicals, Eschwege, Germany); mouse monoclonal anti-glial fibrillary acidic protein (GFAP) (Chemicon, Hampshire, UK); rabbit polyclonal anti-HIF-1α (Cayman Chemical, Ann Arbor, MI); rabbit polyclonal anti-β-tubulin III (anti-TUJ1; Covance, Richmond, CA); and horseradish peroxidase-conjugated secondary antibodies (Pierce, Rockford, IL).

Techniques: Western Blot, Expressing, Confocal Microscopy