Figure 3

Mean expression of genes associated with seven major signaling pathways and validation of expression of key genes using real-time PCR. (A) Supervised cluster analyses of differentially expressed genes (p < 0.05) associated with seven major signaling pathways. The mean expressions of these significant genes are shown among PB, BM and LN cells. (B) Confirmation of the differentially expressed genes among PB-CLL, BM-CLL and LN-CLL using real-time PCR. Expression of the selected genes from each major pathway was studied by using real-time PCR. Significant differences in expression levels between PB-CLL, BM-CLL and LN-CLL were determined by using the Student t test (p values when comparing with LN cells: ^*p < 0.05, ^**p < 0.01, ^#p < 0.005, ^##p < 0.001; p values for comparison with PB is denoted with ^*p and comparison with BM is denoted as ^*b). First row: BCR signaling: SYK, BTK, ZAP70; second row: BAFF/APRIL signaling: BAFF, BCMA, TRAF2; third row: MAPK signaling: MAP2K6, CAMLG, STAT1; fourth row: PI3K/Akt pathway: PDK1, IGFBP6, AKT1; fifth row: NF-κB pathway: NF-κBIB, FCER2, CCND2; sixth row: chemokine ligands/receptors: CXCR4, CCL21, CCR7; seventh row: tolerogenic signature: CAV1, MCM3, IDH; microarray analyses had shown all genes were differentially expressed among PB-CLL (n = 20), BM-CLL (n = 18) and LN-CLL (n = 15).