Figure 1

Expression of cathepsin D and tissue histology in cathepsin D chimaeric mice. a) Leukocytes cells were collected from control unirradiated mice (Balb/c), or from CCDWT or CCDD between 8 and 12 weeks post reconstitution, and stained for CD3 (T cells) or B220 (B cells). The results show the average % of each cell type, from a minimum of 5 different experiments, with standard error of mean. No significant differences (Student's T Test) were observed between any of the three groups of mice. b) Spleens and livers were collected from CCDWT and CCDD mice three-four months post reconstitution, and analysed for cathepsin D and cathepsin E expression by Western blot. The blot for spleen shows the presence of precursor (53KD) and single chain (48KD) forms of cathepsin D. The two single chain degradation products of 31 and 14KD are not shown in this figure. Left panel shows a representative example from one individual mouse from each group. Right panel shows the mean band density (and standard error of the mean) for cathepsin D staining for spleen extracts from five mice taken from three independent experiments. c) Dendritic cells were cultured from bone marrow of CCDWT and CCDD mice, and stained for cathepsin D expression by immunofluorescence. Nuclei are stained with DAPI. At least 100 cells per slide were scored as cathepsin D positive or negative from each sample. The % cells showing cathepsin D staining was greater than 80% for CCDWT, and less than 1% for CCDD mice. d) Spleen from CCDD and CCDWT mice was collected and standard haematoxylin and eosin staining was performed on 4 μm sections of formalin-fixed paraffin embedded tissues. A. Low power view of CCDWT mouse spleen. B. High power view of CCDWT spleen showing PALS (periarteriolar lymphoid sheath) and adjacent red pulp. C. Low power view of CCDD mouse spleen. D. High power view of CCDD mouse spleen showing PALS and adjacent red pulp. e) Upper intestine from CCDD mice was collected and processed as for panel d. Longitudinal and transverse sections show the normal crypt morphology of the adsorptive epithelium.