Fig. 2

CBL0137 induces protective autophagy in B-NHL cells. A Autophagosomes in SU-DHL-4, Raji, and Jeko-1 cells treated with or without 2.0 μM CBL0137 were observed by TEM. The arrow indicates the autophagosome. Scale bars = 2 μm. B The expression of GFP + /mRFP + (yellow) and GFP-/mRFP + (red) LC3 puncta was observed by fluorescence microscopy in B-NHL cells treated with or without 1.0 μM or 2.0 μM CBL0137 for 24 h. Representative images and quantitative analysis of fluorescent LC3 puncta were shown. Scale bars = 30 μm. C Expression of autophagy-related proteins was detected by western blotting in B-NHL cells treated with CBL0137 or DMSO. D–F Effect of chloroquine (CQ) on CBL0137-mediated autophagy, cell viability, and apoptosis. Cells were pretreated with 50 μM CQ for 1 h and then exposed to 1.0 μM CBL0137. D Cell viability was determined by CCK-8. E Apoptosis was measured by flow cytometry. F Conversions of LC3I to LC3II, p62, Atg14, and PARP were examined by western blotting. G B-NHL cells were pretreated with 2.5 mM 3-methyladenine (3-MA) for 1 h and then exposed to 2.0 μM CBL0137. The expression of PARP, p62, LC3, Bcl-xL was detected by western blotting. H 50 μM Z-VAD-FMK was pretreated for 1 h and then exposed to 2.0 μM CBL0137. Caspase-related proteins and autophagy-related proteins were detected by using western blotting