Analysis of LRP and RAP levels in transgenic animals.

<p>Five [APPswe/PS1dE9](+/−)/RAP(+/−) mice (lanes 2, 4, 8, 9), one [APPswe/PS1dE9](−)/RAP(+/+) mouse (lane 1), one [APPswe/PS1dE9](−)/RAP(+/−) mouse (lane 3), one [APPswe/PS1dE9](+/−)RAP(+/+) mouse (lane 5) and two [APPswe/PS1dE9](−)/RAP(−/−) mice (lanes 6, 7) were used in this experiment. 100 µg total protein was loaded per lane. Upper panel - Immunoblot of tissue extract probed with antibody 4109 to RAP (1∶1000). Lower panel - Immunoblot of tissue extract probed with antibody 377 to LRP (1∶1000). The upper band (∼600 kDa) is the full length LRP, the lower band is an 85 kDa fragment of mature, endoproteolytically cleaved, LRP.</p

Genotypes of offsprings between crosses of APPswe/PS1dE9(+)/RAP(+/−) mice and RAP (−/−) mice.

*<p>Both animals died before 3 months of age.</p

Analysis of RAP, LRP and SorLA/LR11 levels.

<p>A). Immunoblots were probed with polyclonal antibody 4109 to RAP (1∶1000), 377 to LRP (1∶1000) and monoclonal antibody anti-LR11 (1∶1000; BD Biosciences, San Jose, CA). Protein concentration was determined by BCA. Each lane contains 100 µg total protein. Genotypes of the mice are marked on the figure. B). Quantification of the intensity of the bands in panel A, using a Fuji LAS-3000 imaging device and software provided by the manufacturer. Statistical comparisons of each protein in the two genotypes of interest ([APPswe/PS1dE9](+/−)/RAP(+/+) and [APPswe/PS1dE9](+/−)/RAP(+/−) were conducted on the raw quantitative data, which consisted of pixel values for each protein band quantified. Two-tailed student t-Test with equal variance was used to estimate the probability that differences in the levels of each protein, between genotypes, resulted from random chance (p values for each comparison are noted on the figure). Because the levels of each protein were measured as lower in mice lacking one RAP allele, we chose to graph the data by setting the mean value for each protein in the [APPswe/PS1dE9](+/−)/RAP(+/+) mice to 100, designated controls, and then graphing the values of mice lacking one RAP allele as a percent of the controls. The data represent measures from 6 [APPswe/PS1dE9](+/−)/RAP(+/+) mice compared to 7 [APPswe/PS1dE9] (+/−)/RAP(+/−) mice (except for measures of LR11 n = 6 as one lane was unmeasurable). The two mice without APPswe/PS1dE9 transgenes were not included in measurements so that the only difference between the two groups of animals analyzed was the number of functional RAP alleles.</p