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. 2021 Jul 19;4(4):1390–1407. doi: 10.1021/acsptsci.1c00091

Figure 3.

Figure 3

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.