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. 2020 Sep 6;6:81. doi: 10.1038/s41420-020-00315-w

Fig. 1. HNK induces autophagosome accumulation, LC3B conversion, and other autophagy related proteins.

Fig. 1

a MCF7 and MDA-MB-231 cells were treated with 5 µM HNK for 24 h and visualized under an electron microscope. Representative pictures are shown with approximately ×7400 magnification (×50000 in the highlighted area). b Double-membrane autophagosomes were counted in randomly selected ~100 cells in random fields. Number of autophagosomes per cell is shown in bar graphs. *P < 0.05, compared with vehicle-treated controls (c) in MCF7 and MDA-MB-231 cells. c Immunoblot analysis of pATG1, ATG5, ATG7, and BECN1 in breast cancer cells treated with 5 µM HNK for indicated time intervals. ACTB was used as a loading control. d Breast cancer cells were treated with 5 µM HNK for indicated time intervals and total cell lysates were immunoblotted for LC3B expression. ACTB was used as a loading control. e Breast cancer cells were treated with 5 µM HNK or 200 nM rapamycin or Earle’s balanced salt solution (EBSS) for 24 h and subjected to immunocytochemistry using LC3B antibody. Scale bars: 20 µm. Representative immunofluorescence images are shown.