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. 2019 Jul 28;9(19):5577–5594. doi: 10.7150/thno.34663

Figure 6.

Figure 6

CPX promotes autophagy flux in CRC cells. A. CRC cells were treated with vehicle, CPX (20 μM), CQ (10 μM), or in combination for 24 hours. Immunoblot analysis was used to detect the expression of LC3 and p62. B. CRC cells were treated with CPX (20 μM) with or without Baf A1 (100 nM) for 24 hours. The expression of LC3 and p62 were examined by immunoblotting. C. CRC cells were treated with E64D (10μg/ml) and PepA (10μg/ml) in the presence or absence of CPX (20 μM) for 24 hours. The expression of LC3 and p62 were examined by immunoblotting. D. The accumulation of LC3 puncta was examined by immunofluorescent analysis of cells treated with CQ (10 μM) in the presence or absence of CPX (20 μM) for 24 hours. Scale bar, 20 μm. The number of LC3 puncta was quantitated. E. Immunofluorescence analysis of cells transiently transfected with tandem mRFP-GFP-tagged LC3 and treated with vehicle, CPX (20 μM), CQ (10 μM), or in combination for 24 hours. Scale bar, 10 μm. The ratio of red puncta indicating autolysosome (GFP-/RFP+) versus yellow puncta indicating autophagosome (GFP+/RFP+) was quantitated. F. Immunofluorescence analysis of the co-localization of endogenous LC3 and LAMP1 in CRC cells treated with vehicle, CPX (20 μM), CQ (10 μM), or in combination for 24 hours. Scale bar, 20 μm. Data are means ± s.d. and are representative of 3 independent experiments. *, P < 0.05, **, P < 0.01, ***, P < 0.001.