Fig. 5: AID−/− B cells and CLL cells have downregulated Smad1/S1pr2 and exhibit less migration in response to S1P.
From: Loss of AID exacerbates the malignant progression of CLL
A A heat map of the top 100 differentially expressed genes from RNA sequencing of CLL cells from 3 Eμ-TCL1 mice and 3 AID−/−/Eμ-TCL1 mice shows that Smad1 and S1pr2 mRNAs are strongly downregulated in AID−/−/Eμ-TCL1 CLL cells compared to Eμ-TCL1 CLL cells. B, C B cells and CLL cells were examined for the expression of S1PR2 (B) and S1PR1 (C) in the spleens of 4-month-old Eμ-TCL1 (n = 9) and AID−/−/Eμ-TCL1 (n = 9) mice by flow cytometry. D Transwell assay of migration of B cells and CLL cells purified from the spleens of Eμ-TCL1 (n = 4) and AID−/−/Eμ-TCL1 (n = 4) mice towards 10 nM S1P. E B cells and CLL cells purified from the spleens of Eμ-TCL1 (n = 4) and AID−/−/Eμ-TCL1 (n = 4) mice were pretreated with 10 μM of the S1PR2 inhibitor JTE-013 for 30 min before the transwell migration assay towards 10 nM S1P. F B cells purified from the spleens of WT and AID−/− mice were stimulated with 20 μg/mL LPS for 3 days and lysates were immunoblotted for the indicated proteins. G B cells purified from the spleens of WT and AID−/− mice were stimulated with 20 μg/mL LPS for 3 days and examined for the expression of S1PR2 by flow cytometry.
