Figure 5.

Analysis of myelin markers. A, MBP Western blot analysis of total brain homogenates (50 μg/lane) from FA2H^+/+, FA2H^+/−, and FA2H^−/− mice of different age groups. Membranes were stained with anti-MBP antiserum recognizing all MBP isoforms and anti-actin antibody to control for equal loading. B, Bound antibodies were detected by enhanced chemiluminescence. Densitometric quantification of MBP levels. Linear exposures of x-ray films were scanned, and the intensities of FA2H^+/+ were set to 100%. Shown are the mean ± SD (n = 6 for P10; n = 3 for P19–P70). No significant differences between the three genotypes were observed. C, Western blot analysis of CNP. Brain homogenates of the indicated genotypes and ages were stained with anti-CNP antibody and anti-actin antibody as loading control. D, Western blot analysis of L-MAG and NCAM in 70-d-old brain homogenates. Relative protein levels (normalized to actin) of L-MAG and NCAM-120 are shown on the right (n = 3). E, Ratio of S-MAG and L-MAG expression levels at P10 and P28 was determined by RT-PCR. Intensities of ethidium bromide-stained PCR products were measured (n = 3). F, Quantitative real-time RT-PCR of MBP, PLP, and MAL expression (normalized to actin expression) at P10 and P28 and in adult FA2H^−/− and FA2H^+/+ mouse brains (12 weeks) showed upregulation of these myelin genes during the period of myelination, as expected. However, no statistically significant differences in their expression levels were observed at any time point examined (mean ± SEM; n = 3–5).