Figure 1
(a) Whole-brain iron levels by inductively coupled plasma atomic emission spectroscopy (ICP-AES). Mean±s.e.m., Student’s t-test *P=0.002, n ≥5/group. (b and c) Immunoblot analysis of ferritin relative to β-actin. Mean±s.e.m., Student’s t-test *P=0.0005, n ≥5 per group. (d) Real-time reverse transcription-PCR (RT-PCR) validation of changes in transcript levels for selected neurodegeneration with brain iron accumulation (NBIA) genes. Mean±s.e.m., Student’s t-test *P<0.04, n=7/group. (e) Luxol Fast Blue staining (left panels) was analyzed by the Positive Pixel Count algorithm (right panels) that grades areas as high (red), moderate (orange) or weak (yellow) positive or negative (blue) staining. (f and g) Average density (total intensity of all positive pixels divided by total number of positively stained pixels) and positivity (number of positive pixels divided by the total pixels; measures proportion of brain area staining positive) of Luxol Fast Blue staining for myelin were unchanged. Mean±s.e.m., Mann–Whitney test P>0.05, n≥4/group. (h) Electron micrographs of myelinated axons (top) and glial cell organelles (bottom) in the corpus callosum; oligodendrocyte mitochondria (red arrows) and Golgi apparatus (yellow stars). (i and j) Activity measures (number of visits and duration of visits per test group per day) in the habituation (Habit.) and nose-poke adaptation (NPA) phases of IntelliCage testing. Mean±s.e.m., two-way ANOVA (i) *P<0.0112 (Habit.) and **P<0.0004 (NPA); (j) *P<0.0007 (Habit.) and **P<0.0001 (NPA), n ≥6/group.
