Fig. 3: Migration of EBV-infected B cells is dependent on FAK2.

a EBV-infected B cells were exposed to the FAK2 inhibitor defactinib (Def.) for 30 min (3.5 µM) and seeded in collagen at a concentration of 3 × 10E5 cells/ml. 2D cell tracks with or without alignment were generated from time lapse microscopy recordings. The bar graphs show the mean percentage of motile cells with standard deviation in treated and untreated cells (n = 3 independent B cell samples). Treated EBV-infected B cells were also observed by scanning electron microscopy. Curves showing EBV-infected cell growth in the presence of defactinib (Def.) at a 3.5 µM concentration over two weeks. Mean cell count and standard deviation are indicated (n = 5 independent B cell samples). b Total and phosphoFAK2 expression levels in B cells stimulated with CD40L, IL-4 and CXCL12 or after EBV infection were determined by western blot with specific antibodies. The bar graph summarizes the mean phosphoFAK2 expression levels and standard deviation, relative to the loading control tubulin (n = 4 independent B cell samples). c Western blot showing phosphoFAK2 expression levels in EBV-infected B cells exposed to the CCR1 inhibitor BX471 at 10 μM for 30 min. The arrow refers to phosphoFAK2, the asterisk refers to a non-specific signal. The bar graph summarizes the mean phosphoFAK2 expression levels and standard deviation. Values are given relative to the values observed in the untreated population. An immunoblot against tubulin was used as loading control (n = 4 independent B cell samples). d Curves showing EBV-infected cell growth over time in the presence of defactinib (Def.) at a 0.5 µM concentration. Infected cells were alternatively treated with BX471 (5 µM) alone or with a combination of low doses defactinib (0.5 µM) and BX471 (5 µM). Mean cell count and standard deviation are indicated (n = 5 independent B cell samples). e dose-response curve for defactinib and BX471 (n = 3 independent B cell samples, each point showing mean and standard deviation), together with isobolograms (ED₁₀ = 10% of maximal effect and ED₃₀ = 30% of maximal effect). The linear curve shows additivity of the drug effects, the curve below it is indicative of synergistic effects. Statistical significance was determined using two-sided paired t tests in (a) (growth curve at day 14), b and (c) and one-way analysis of variance at day 7 in (d). *p < 0.05, **p < 0.01. Source data are provided as a Source Data file.