Figure 1
Autophagy activation by nPt in HchEpC1b cells, an EVT cell line. (a) Western blots of HchEpC1b cells, which were cultured with 25 μg/ml of nPt for 24 h with or without E64d (E64) and pepstatin A (P) for 2 h at the end of culture, were as follows: MAP1LC3B (LC3), SQSTM1, and ACTB. The expression levels of MAP1LC3B-II (b) or SQSTM1 (c) in the HchEpC1b cells cultured with nPt were shown in the graphs. Expression was normalized with ACTB levels. (d) Representative panels showed the merged images of anti-MAP1LC3B staining (LC3, green) and nuclei staining (DAPI, blue) of HchEpC1b cells cultured with 25 μg/ml of nPt for 24 h in the presence or absence of E64d and pepstatin for 2 h. (e) The graph showed the average number of LC3 puncta in HchEpC1b cells treated with the presence (black bars) or absence (white bars) of E64 and P, as shown in (d). (f) The cell number of HchEpC1b cells with nPt at the indicated concentrations (μg/ml) for 24 h were shown. The numbers of cells in the treatment groups were normalized to that with control treatment, PBS, as one. Data were expressed as the mean ± S.D. *p < 0.05, **p < 0.01, ***p < 0.001. Scale bars: 20 μm.
