Supplemental figure 2. PIM kinases are a potential therapeutic target in neuroblastoma from PIM Kinases Are a Potential Prognostic Biomarker and Therapeutic Target in Neuroblastoma

Abstract

<p>(A-B) Knockdown levels of PIM1, -2, and -3. KELLY and SH-SY5Y cell lines were transfected with scrambled siRNA (siCTRL) or siRNAs targeting PIM1, -2, or -3, cultured for 2 days, followed by RNA isolation and qRT (n=3). (C-D) Neuroblastoma cell lines demonstrate differential sensitivity to AZD1208 in a short-term assay. Cells were plated in 384-well plates, treated for 5 days with increasing concentrations of AZD1208, and viability was measured using CellTiter-Blue (n=3). (E-F) Neuroblastoma cell lines demonstrate differential sensitivity to PIM-447 in a colony formation assay. Cells were plated in 6-well plates and treated with increasing concentrations of AZD1208. Drug-containing medium was refreshed every 3 days. Cells were fixed, stained, and scanned when untreated wells reached confluency (n=3). (G) PIM-447-resistant cell lines demonstrate sustained mTOR signaling upon treatment. Cells were treated with 2 μM PIM-447 for 0, 24, and 48 hours followed by western blot analysis (n=3).</p>