FOXO1 knockdown is toxic for BCP-ALL cell lines

We found that FOXO1 knock-down inhibits cell cycle progression and induces apoptosis in BCP-ALL cell lines RS4;11 and 018Z. BCP-ALL cell lines RS4;11 and 018Z were transduced with sh_hFOXO1.H-10381 in pRSI12-U6-sh-UbiC-TagRFP-2A-Puro lentiviral vector (DECIPHER shRNA Library, Human Module I (Cellecta, https://www.cellecta.com/) obtained from BioCat, Heidelberg, Germany). On Day 5 after transduction total RNA was isolated with RNeasy mini kit (QIAGEN). Microarray analyses were performed using 200 ng of total RNA as starting material and 5.5 µg ssDNA per hybridization (GeneChip Fluidics Station 450; Affymetrix, Santa Clara, CA). The total RNAs were amplified and labeled following the Whole Transcript (WT) Sense Target Labeling Assay (http://www.affymetrix.com). Labeled ssDNA was hybridized to Human Gene 1.0 ST Affymetrix GeneChip arrays (Affymetrix, Santa Clara, CA). The chips were scanned with a Affymetrix GeneChip Scanner 3000 and subsequent images analyzed using Affymetrix® Expression Console Software (Affymetrix). Probe level data were obtained using the robust multichip average (RMA) normalization algorithm.