Figure 7.

Chloroquine significantly reduced the efficiency of adipogenesis in primary MEFs. Wild-type primary MEFs were induced for adipogenesis with or without co-treatment of 10 μM chloroquine (CQ). Differentiation progress was then monitored by: (A) microscopy analysis; (B) lipid analysis with Bodipy 493/503 staining (14 days after differentiation induction); (C) lipid analysis by spectrometry of Oil Red-O staining (14 days after differentiation induction). (D and E) are controls that show that chloroquine was nontoxic (D) and efficacious in inhibiting autophagosome fusion with lysosome and in inhibiting autophagy flux (E) at the experimental concentration. (D) Tunel assays of wild-type MEFs treated with 10 μM chloroquine for 4 days compared with cells without chloroquine treatment. Cells treated with 10 μM staurosporine (STS) for 6 hr was used as a positive control. (E) Cells treated with/or without 10 μM chloroquine at different time points were harvested, immunoblotting assays were performed with LC3, p62 or RAN antibodies, as indicated. The levels of RAN served as a loading control. The results represent three independent experiments. *p < 0.05; Student t-test.