Curcumin stimulates autophagy initiation. (a) Formation of GFP-LC3 puncta in HEK293 cells stably expressing GFP-LC3. The cells were treated with curcumin, rapamycin or chloroquine (CQ), a lysosome inhibitor, respectively. Scale bars, 10 µm. (b) Statistical analysis of the number of GFP-LC3 puncta per cell in (a). Data are shown as mean ± SEM; n = 30. ***P < 0.001. (c) The protein level of p62 in WT MEFs or Atg7−/- MEFs treated with curcumin, rapamycin or chloroquine, respectively. (d) Quantification of p62 protein level in (c). The statistical data are presented as mean ± SEM of three independent experiments. ***P < 0.001. (e) Formation of GFP-DFCP1 puncta in HEK293 cells stably expressing GFP-DFCP1. The cells were treated with curcumin or rapamycin. Scale bars, 10 µm. (f) Statistical analysis of the number of GFP-DFCP1 puncta per cell in (e). Data are shown as mean ± SEM; n = 30. **P < 0.001