Fig. 8
From: Critical and differential roles of eIF4A1 and eIF4A2 in B-cell development and function
eIF4A1 exerts its functions in B cells through Gins4 and other genes. A Immunoblot analysis of Gins4 and eIF4A1 protein expression in LPS-stimulated Cas9-GFP B cells transduced with retroviruses encoding nontargeting control (NTC), Eif4a1 or Gins4 sgRNA. The retroviral vector encodes blue fluorescent protein (BFP), and transduced cells are BFP positive. B Cas9-GFP and wild-type B cells (CD45.1 + ) were mixed at a 1:1 ratio, stimulated with LPS for 24 hours, transduced with retroviruses encoding nontargeting control (NTC), Eif4a1 or Gins4 sgRNA, and analyzed by flow cytometry at 72 hours post retroviral transduction. The proliferation ratio was calculated as indicated. C Scheme of iGCB culture. D Immunoblot analysis of eIF4A1, eIF4A2 and Gins4 proteins in Eif4a1fl/fl and Eif4a1fl/fl;Cγ1Cre B cells cultured in the iGCB system at the indicated time points. E, F Numbers of Eif4a1fl/fl and Eif4a1fl/fl;Cγ1Cre iGCB E and iPC F cells at the end of the indicated stages of culture. G Eif4a1fl/fl and Eif4a1fl/fl;Cγ1Cre B cells were cultured in the iGCB system and transduced with retroviruses encoding Eif4a1 or Gins4 at iGCB day 2.5. Transduced cells (GFP + ) were sorted at iGCB day 4 and replated on 40LB cells in the presence of IL-21 for iPC differentiation. The numbers of cells at the end of iPC culture were plotted. The data shown are representative of three independent experiments
