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Table 1.

Number of up and down regulated genes.

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Table 2.

Overlapping genes between different treatments.

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Table 3.

Antigen expression determined by FACS analysis.

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Table 4.

Antigen expression determined by Affymetrix analysis: mean expression values of untreated cells.

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Figure 1.

Comparison of genes deregulated by different antibody treatments.

Venn diagrams for numbers of up- (A) or down- (B) regulated genes in SUDHL4 after a 4 h incubation with 10 µg/ml anti-BCR antibody (anti-IgG) and anti-CD20 antibodies (Rituximab, LT20) and Venn diagrams for up- (C) or down- (D) regulated genes in REC1 after treatment with anti-BCR antibodies (anti-IgG and anti-IgM, respectively) and anti-CD20 antibodies (Rituximab, Rx-F(ab')2, LT20).

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Figure 2.

Comparison of anti-BCR and anti-CD20.

Venn diagrams of “Union lists” of up- (A) or down- (B) regulated genes after different treatments. Cells were incubated with 10 µg/ml antibody for 4 h. Lists of similar treatments were united to “Union lists” for CD20 or BCR treatment for each cell line R = REC1, S = SUDHL4.

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Figure 3.

Comparison of anti-BCR treatments in all cell lines studied.

Venn diagrams of “Union lists” ( = Union of lists for similar treatments) for anti-BCR treatment of up- (A) or down- (B) regulated genes in OciLy18, Z138, SUDHL4 and REC1.

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Table 5.

Functional clustering of genes deregulated by anti-BCR antibodies in cell lines OciLy18, Z138, REC1 und SUDHL4.

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Table 6.

Functional clustering of genes deregulated by anti-CD20 antibodies in SUDHL4 cells und REC1 cells (KEGG-, Panther- und Biocarta, GO terms).

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Table 7.

Genes that are common deregulated in all four cell lines by anti-BCR antibodies and in SUDHL4 and REC1 by anti-CD20 antibodies.

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Figure 4.

Interactions between deregulated genes.

IPA generated network of interactions between genes deregulated by anti-CD20 antibodies in SUDHL4 and REC1. The red circle marks genes which are also deregulated by all anti-BCR treatments. Interestingly this unbiased analysis displays deregulations of BCR components and the downstream signaling pathway.

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Figure 5.

Repression of chemokine CCL3/4 induction caused by Syk inhibition and silencing.

(A) CCL3 secretion in SUDHL4 induced by anti-IgG antibodies and anti-CD20 antibodies and its inhibition by Syk inhibitors I and IV (Calbiochem) at 1 µM and 0,32 µM respectively. Bars represent mean of three replicates, including standard deviation (<IgG> = anti-IgG antibody, Inh. I = Syk inhibitor I, Inh. IV = Syk inhibitor IV, LT20 = murine anti-CD20 antibody, Rx-Fab'2 = F(ab')2 fragment of rituximab). Inhibition of CCL3 (B) and CCL4 (C) secretion in SUDHL4 induced by anti-IgG antibodies or rituximab after siRNA-mediated Syk silencing. Results represent at least three independent experiments. Students t-test for “Negative controls” vs. “SYK-knockdown”: for CCL3 secretion after anti-IgG treatment p = 6.8*10−7, Rx treatment p = 7.4*10−3. For CCL4 secretion after anti-IgG treatment p = 2.1*10−6, Rx treatment p = 2.5*10−4. (anti-IgG = anti-IgG antibody, Rx = rituximab, Neg. controls = RISCfree and Luciferase-siRNA transfections, respectively; SYK knock down = Syk-siRNA transfections).

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