Figure 5
From: Haematopoietic development and immunological function in the absence of cathepsin D
a) Electron microscope images of spleen cells from CCDD (A-C) and CCDWT (D). Macrophages (one example marked with asterisk in panel A) in the marginal region of the white pulp contain abundant lysosomal/vacuolar structures, most of which resemble granular osmiophilic deposits (B, a higher power of the quadrated area in A). Lymphocytes in the region appear intact (A). In the red pulp, such macrophages with abundant lysosomal structures (arrow) appear to be less in number (C). Macrophages (asterisk) and lymphocytes in the spleen of CCDWT show normal structures (D). b) Western blot showing expression of LC3 and cathepsin E (as loading control) in spleens from CCDWT and CCDD. A blot of CCDWT liver with LC3 antibody is shown for comparison. Arrows show the position of the soluble (LC3I – 18kD) and membrane bound (LC3II – 16kD) forms of LC3. c) Distribution of LC3 in spleen cells from CCDD (LC3 was not detectable above background in CCDWT). Left panel – LC3 staining. Right panel – autofluorescence. Note presence of high levels of LC3 in a cell with strong autofluorescent granules (arrow). Lower panels shows autofluorescent granules in controls, without primary antibody.