Figure 7.

Modulation of autophagy affected the cytotoxicity effect of leflunomide in 5637 cells. Combination use of leflunomide with 10nM rapamycin or 20nM CQ for 48 hours led to increased or reduced, respectively, autophagy levels compared with the single use of leflunomide, as determined by Acridine Orange staining (A) and Western blot (B and C). Microscopy observation (D) showed an inferior cell viability in the presence of leflunomide and chloroquine, compared with leflunomide alone. By contrast, the restored cell abundance and morphology was observed with the combined use of leflunomide and rapamycin. MTS assay (E), cell apoptosis analysis by flow cytometry (F and G) and Western blot (H and I) exhibited consistent results. Beta-actin or Beta-tubulin was used as internal loading reference. The results (C, E, G and I) represent the mean ± SD from three independent experiments. *P<0.05; **P<0.01.

Abbreviation: CQ, chloroquine.