Figure 3. Altered dynamin staining in glomerulus after LPS treatment is CatL dependent.

(A) Immunogold analysis of dynamin in podocyte FPs of rat glomerulus. Low-power view (left) depicts dynamin within the center of podocyte FPs (asterisk) as well as in electron-dense areas rich of cortical actin (a) (original magnification, ×30,000). High-power view shows gold particles, which are associated with the cytoplasmic side of vesicles (v) and with electron-dense actin areas (a) (original magnification, ×45,000). E, endosome; GBM, glomerular basement membrane; Ly, lysosome; SD, slit diaphragm; US, urinary space. (B) Immunocytochemistry of dynamin in glomeruli of WT mice before and after injection of LPS and of CatL^–/– mice after injection of LPS. (C) Immunocytochemistry of samples from CatL^–/– mice for glomerular dynamin and CatL. CatL^–/– mice had been reconstituted with either Pre-Pro-CatL (left) or with short CatL (right) using transient gene transfer. Twenty-four hours after gene transfer, glomeruli were stained using anti-CatL and anti-dynamin hudy 1 antibody. Original magnification, ×400 (B and C). (D) Immunoblot of endogenous dynamin in cultured podocytes in response to LPS. Recombinant dynamin (REC), extracts of WT and CatL^–/– podocytes, were analyzed. (E) Quantitation of dynamin in cell extracts shown in D. Dynamin signal was adjusted to actin levels.