Evaluation of S14161 analogues. (A) The chemical structures of S14161, S14147, and compounds 1-4 (CMP1, CMP2, CMP3, and CMP4). (B) Starved KMS11 cells were treated by S14161, its analogs, or DMSO for 2 hours followed by 100 ng/mL IGF1 for 10 minutes. After incubation, cells were harvested and total proteins isolated. Expression of AKT, phospho-AKT (p-AKT), and β-actin was measured by immunoblotting. LY indicates LY294002. (C) KMS11 cells were treated for 24 hours with compounds 1-4 or S14147 or vehicle DMSO (each at 10μM). After incubation, cells were harvested and total proteins isolated. Expression of cyclin D2 (CCND2) and β-actin was measured by immunoblotting. (D) JJN3 cells were treated with 10μM S14161, S14147, compounds 1-4, or buffer control for 24 hours. After incubation, apoptosis was measured by annexin V–fluorescein isothiocyanate (FITC) and propidium iodide staining. A representative experiment is shown.