Fig. 4

FBXO17 depletion reduces cell proliferation and dysregulates the PI3K/Akt/mTOR pathway. a A549 cells were transfected with scrambled siRNA or siFBXO17. Live cells were counted 96 h after transfection using trypan blue exclusion staining. Data shown is representative of three independent experiments, **p < 0.001. b A549 cells were transfected with scrambled siRNA or empty vector. After 48 h cells were plated at 5 × 10⁴/well in a 96-well plate. The MTS reagent was added at 24, 48, and 72 h after seeding. Following 30 min of substrate addition samples were analyzed by spectrophotometry in triplicate. Data shown is representative of three independent experiments, *p < 0.05, **p < 0.01, ***p < 0.001. c After 72 h post transfection of scrambled siRNAs or siFBXO17, A549 cells were harvested in triplicates and RNA was isolated for analysis using the Clariom S RNA microarray. Using cutoffs of +/− 1.5-fold change a volcano plot was created representing 212 genes (Transcriptome Analysis Console Software, version 4.0.1). d Pathway analysis of Clariom S transcriptome profiling using ToppGene revealed decreased expression of nine genes involved in cell proliferation and metabolism of tumor cell lines (consistency score 2.667). e Immunoblot of CKS1B is shown in control (scrb) and sifbxo17 cell lysates. f ToppGene was used to analyze the gene sets and determine enrichment of biological pathways and diseases