Additional file 6: of Small Molecule KRAS Agonist for Mutant KRAS Cancer Therapy

Figure S6. Silencing of mutant KRAS reduced sensitivity of cells to KRA-533. (A) KRAS shRNA plasmids were transfected into A549 cells that contain KRAS mutation, followed by Western blot using KRAS antibody. (B) A549 cells and mutant KRAS silenced A549 cells were treated with KRA-533 (15 μM) for 48 h. Apoptotic cells were detected by Annexin V /PI binding and analyzed by FACS. Data represent mean ± SD, **P < 0.01, by 2-tailed t test. (C) GFP-LC3 constructs and KRAS shRNA plasmids were co-transfected into A549 cells, followed by treatment with KRA-533 for 48 h. Autophagic cells (GFP-LC3 positive cells) were visualized by Axioplan Zeiss microscope and quantified. Data represent mean ± SD, **P < 0.01, by 2-tailed t test. (JPG 653 kb)