S14161 inhibited phospho-AKT translocation and accumulation at the cytoplasmic membrane. OPM2 myeloma cells were starved overnight, followed by treatment with S14161 (100μM for 1 hour), LY294002 (100μM for 30 minutes), or DMSO control for 1 hour. Cells were then treated with 100 ng/mL IGF1 for 10 minutes. Cells were fixed and stained with antibodies against AKT or phospho-AKT (p-AKT) and DAPI (4,6 diamidino-2-phenylindole) as described in “Immunofluorescence assay.” Red indicates AKT or p-AKT; blue, nuclei. (A-C) Phospho-AKT (arrows) stimulated by IGF; (D-F) total AKT translocated to the plasma membrane after IGF1 treatment; (G-I) Phospho-AKT inhibited by LY294002; (J-L) total AKT located in the nuclei after LY294002 treatment; (M-O) phospho-AKT inhibited by S14161; and (P-R) total AKT was restricted to the nuclei after S14161 treatment.