Figure 1

Constructs and screening system. (A) 293FT/RFP1 cells stably express RFP1 fluorescent protein. RFP1 florescence loss after AID expression is used to assess AID mutagenic activity. (B) Schematic representation of the AID expression vector. A CMV promotor is followed by a human AID or AID mutant gene which is linked to an HA tag at the C-terminus followed by furin/2 A peptide (F2A) bicistronic expression linker and an eGFP reporter gene. An internal ribosome entry site (IRES) is used for bicistronic expression of a puromycin resistance gene. (C) Cell lysates from 293FT/RFP1 cells expressing AID mutants were used to perform immunoblot analysis with antibodies binding to the HA tag on AID or tubulin as a cell loading control.