Figure 7: Functional characterization of cell dynamic changes in Eμ-NKX2-3 transgenic mice.

(a) Migration frequencies of transgenic B cells non-treated (none) or treated with the indicated chemical inhibitors (AMD, CXCR4 antagonist, 10 μM; A286, LFA-1–ICAM-1 interaction inhibitor, 10 μM; anti-LFA1, blocking antibody anti-mouse CD11a, 1 μg ml⁻¹) in Boyden chambers in the absence (none-chemokine) or presence of CXCL12 (10 nM). Significance levels of changes are indicated (two-tailed Student’s t-test: *P<0.05, **P<0.001, ***P<0.0001) in all a–f cases. (b) Frequency of adhesion to ICAM-1-containing artificial membranes of transgenic B cells none-treated or treated with the indicated chemical inhibitors. (c) Cell polarization frequencies, indicating the fractions of non-motile and motile cells, of transgenic B cells none-treated or treated with the indicated chemical inhibitors and settled on ICAM-1-membranes in the absence of CXCL12 chemokine. Data in a,b,c are the mean±s.d. (n=4). (d) As in a using the chemical inhibitors Dasatinib (Das, 1 μM; Lyn inhibitor) and P505-15 (P505, 1 μM; Syk inhibitor). (e) As in b using the indicated inhibitors. (f) As in c using the indicated inhibitors. Data in d,e,f are the mean±s.d. (n=3). (g) Survival of splenic Eμ-NKX2-3 CD19⁺ B-cell lymphoma cell cultures incubated during 24 h with the BTK inhibitor (Ibrutinib), the PI3K inhibitor (Idelalisib), the MALT1 proteolytic inhibitor (MI2), the NF-kB inhibitor (Bay11-7082), bendamustine and fludarabine at increasing concentrations. Non-treated cells (NT) were incubated with dimethylsulphoxide (DMSO). IC₅₀, half maximal inhibitory concentration. Error bars indicates s.d.