C8 inhibition of native untagged AID and endogenous AID from B
lymphoma cells. (A) Representative alkaline cleavage experiment demonstrating
C8 inhibition of untagged native AID in 293T whole cell lysate (left
panel). Catalytic activity of eukaryotic-expressed native untagged
AID as a function of log C8 concentration, using 140 mM DMSO as control
for 100% AID activity (right panel). (B) Deam-specific PCR was used
to detect activity of endogenous AID in extracts of AID-expressing
B lymphoma cell lines, with or without addition of C8. The deamination
substrate plasmid was incubated with cell extracts containing endogenously
expressed AID from Raji (top panel), Ramos (middle panel), and Daudi
(bottom panel) with added vehicle (140 mM DMSO) or C8. Incubation
with C8, but not DMSO, abrogated any detectable PCR product. The bottom
gel is a representative experiment demonstrating inhibition of AID-His
using Deam-PCR. (C) Deam-PCR was used to demonstrate inhibition of
AID expressed in 293T cells, in whole cell extract incubated with
the target plasmid.