Differential carbonic anhydrase activities control EBV-induced B-cell transformation and lytic cycle reactivation
Fig 5
Carbonic anhydrase inhibitor retards EBV induced B-cell transformation.
(A-B) PBMCs isolated from two individual donors (donor #1 and donor #2) were infected with GFP-EBV (MOI ~10) in the absence (DMSO control) or in the presence of 0.1 mM carbonic anhydrase inhibitor S4 for 28 days. At the indicated time points post-infected cells were photographed using a Fluorescent Cell Imager. Scale bars, 100 μm. (C-D) In a similar experimental set up, cells at the indicated time points were harvested and subjected to either (C) carbonic anhydrase activity assay, (D) intracellular pH detection and (E) qPCR analyses. (C) Carbonic anhydrase activity and (D) intracellular pH determination assays were performed using kits as per manufacturer’s protocols. (E) The relative changes in transcripts (log10) of the selected genes using the 2-ΔΔCt method are represented as bar diagrams in comparison to uninfected PBMC-control using B2M as housekeeping gene. (C-E) Average values +/- SEM are plotted of two independent experiments performed in similar settings. *, **, *** = p-value < 0.01, 0.005 and 0.001 respectively. (F) Reanalyses of RNA-Seq data (GSE125974 and DRA011328) of B-cells infected with EBV for the indicated time points showing temporal expression prolife of CA9 transcript.