Fig. 4

CPI-613 promotes the AMPK-ULK1 autophagic signaling in pancreatic cancer cells. a, b The effects of CPI-613 on AMPK-dependent autophagic signaling. AsPC-1 and PANC-1 cells growing in 2D culture dishes (a) or the SeedEZ 3D scaffold (b) were treated with or without 200 μM CPI-613 for 48 h, followed by Western blot with the indicated antibodies. c, d The effects of CPI-613 on autophagy. AsPC-1 and PANC-1 cells were transfected with mRFP-GFP-LC3B plasmid and treated with or without 200 μM CPI-613 for 48 h, followed by fluorescence analysis and quantification of LC3B-positive autophagosomes (c). d The effects of CPI-613 on autophagy determined by TEM. AsPC-1 and PANC-1 cells were treated with or without 200 μM CPI-613 for 48 h and subjected to TEM analysis. Autophagosomes in CPI-613-treated AsPC-1 cells were indicated in boxes. e The effects of ULK1 knockdown (shULK1) on LC3B-II under the influence of CPI-613. **p < 0.01