Fig. 3

EZN4150 inhibits p110α and Vps34. a, b SKBR3 green fluorescent protein (GFP)-LC3 cells were transfected with 50 nM control non-targeting siRNA (siCTL) or two siRNA sequences targeting PIK3CA (si38 and si39) and cultured for 4 days. Parallel plates were cultured for 4 days with dimethyl sulfoxide (DMSO) or 500 nM BYL719. Cells were treated with DMSO control (D) or 1 μM Lapatinib (L) for the final 18 h. a Lysates of cells were evaluated by immunoblot analysis with the indicated antibodies. b GFP-LC3 localization was captured by fluorescent microscopy. c Alignment of the EZN4150 sequence with the sequences of human PIK3CA and PIK3C3, the genes encoding p110α and Vps34, respectively. Misaligned bases are underlined in bold. d Cells were treated with 5 μM EZN3046 or EZN4150 for 7–8 days. Lysates were analyzed by immunoblot with the indicated antibodies