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. 2009 Dec 28;107(4):1630–1635. doi: 10.1073/pnas.0908953107

Fig. 5.

Fig. 5.

γ-Secretase inhibition using L685,458 induces the AD-like endosomal phenotype in 2N fibroblasts. (A) Aβ-40 and Aβ-42 levels in cell-culture media collected 18 h after incubation with 10 μM of L685,458 were determined by ELISA (*, P < 0.05; **P < 0.01) . (B) Western blot analysis of cell lysates probed by C1/6.1 for APP and CTFs and by 6E10 for βCTF. Immunolabeling showed EEA1-positive vesicles in (C) DMSO- and (D) L685,458-treated fibroblasts. (E) The relative number of endosomes of different size ranges (mean ± SEM, n = 90; **, P < 0.01; ***, P < 0.001) in DMSO- and L685,458-treated 2N fibroblasts. (F) The relative number of endosomes of different size ranges (mean ± SEM, n = 74; **, P < 0.01; ***, P < 0.001) in shAPP-transfected and untransfected 2N fibroblasts. Western blot analysis (G) and its quantitation (H) of transferrin uptake in L685,458-treated versus control 2N fibroblasts normalized to GAPDH at 5 and 20 min. **, P < 0.01.