FIG 7.

Acetylation-defective mutants of ADA3 fail to rescue the block in cell proliferation imposed by Ada3 deletion in Ada3^FL/FL MEFs. Ada3^FL/FL MEFs stably expressing either empty vector, ADA3 WT, or acetylation-defective mutants were infected either with GFP only or with GFP-Cre expressing adenovirus, and then CellTiter-Glo, Western blotting, or colony formation assays were performed in these cells. (A to E) Relative luminescence units of various Ada3^FL/FL MEFs at various days after the infection are shown. The RLU shown here are means ± SD from three independent experiments, each done in three replicates, and P values were computed using Student's t test. (F to J) Western blots showing the expression of mADA3 and hADA3 WT or mutant proteins. HSC70 was used as a loading control. (K) A composite computation of percent rescue with respect to day 1. Error bars show means ± SEM from three independent experiments (*, P < 0.05; **, P < 0.01, computed using Student's t test). (L to N) Colony formation assay was performed 7 days after the infection, and percent rescue was calculated as described in Materials and Methods. Expression of mADA3 and hADA3 WT or mutants was examined by Western blotting with HSC70 as a loading control.