Figure 7.
Figure 7. Nuclear destabilization of AID is not restricted to B cells. (A) Human 293T cells transfected with WT-AID-HA–expressing vector were incubated for 6 h with both LMB and MG132, or left untreated, as indicated. Immunoprecipitation was performed using agarose-conjugated anti-HA antibodies, and SDS-PAGE–fractionated protein extracts were analyzed by Western blot with anti-AID antibodies. Migration position of AID-HA is indicated. (B) 293T cells stably transfected with a WT-AID-EGFP–expressing vector were incubated with cycloheximide (CHX; 20 or 50 μg/ml) and LMB, either separately or in combination, and the decay of the protein was followed by the decrease in fluorescence intensity. The percentage of initial fluorescence is plotted at various time points after addition of the inhibitors of protein synthesis and/or protein nuclear export.

Nuclear destabilization of AID is not restricted to B cells. (A) Human 293T cells transfected with WT-AID-HA–expressing vector were incubated for 6 h with both LMB and MG132, or left untreated, as indicated. Immunoprecipitation was performed using agarose-conjugated anti-HA antibodies, and SDS-PAGE–fractionated protein extracts were analyzed by Western blot with anti-AID antibodies. Migration position of AID-HA is indicated. (B) 293T cells stably transfected with a WT-AID-EGFP–expressing vector were incubated with cycloheximide (CHX; 20 or 50 μg/ml) and LMB, either separately or in combination, and the decay of the protein was followed by the decrease in fluorescence intensity. The percentage of initial fluorescence is plotted at various time points after addition of the inhibitors of protein synthesis and/or protein nuclear export.

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