Figure 3

Gene expression analysis of BMSCs co-cultured in direct contact with leukemia cells shows changes in IL-17 signaling-related genes. BMSCs from three healthy donors were co-cultured in direct contact with the three different leukemia cell lines (TF-1, TF-1α and K562). The cells were harvested at 4 h, 10 h and 24 h and total RNA were extracted. The total RNA from BMSC mono-cultures was mixed with the total RNA from TF-1, TF-1α or K562 cell mono-cultures and the resulting three mixed total RNA samples were used as a “mono-culture” control and were subjected to gene expression analysis. The RNA from BMSCs and leukemia cells co-cultured in direct contact was extracted and gene expression was analyzed. Analysis with Partek Genomic Suite revealed that 544 genes that were differently expressed between the BMSCs and leukemia cell direct contact co-culture samples and the mono-culture controls (p-value < 0.05, FDR < 0.01). Panel A; Hierarchical clustering analysis of 544 differentially expressed genes in BMSCs and leukemia cell direct contact co-cultre samples compared with the mono-cultures (control) using Partek Genomic Suite program (ANOVA test with unadjusted p-value < 0.05). The displayed colors represent the fold changes where shades of red and blue indicate up- and down-regulation respectively. The color key for the sample labels is on the top left. Panel B. Ingenuity Pathway Analysis (IPA) of the 544 differentially expressed genes. Numerical symbols at the right side of each bar indicate the total number of genes composing the pathway. The bars indicate the percentage of up- (red bar) and down-regulated (green bar) genes in each pathway, while the orange line indicates minus-log transformed p-value. The top 10 canonical pathways are shown.