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. 2019 Dec 24;19:683–695. doi: 10.1016/j.omtn.2019.12.015

Figure 3.

Figure 3

APRT mRNA and Enzymatic Analyses

(A–C) APRT mRNA levels of mutants S23 (A), S62 (B), and S1 (C) and their repaired counterparts obtained after transfection with the corresponding repair-PPRHs and subsequent selection are shown. In (C), also represented are the aprt mRNA levels in three clones repaired using the long-distance repair-PPRH (LD-HpS1E2rep). APRT mRNA levels were determined by qRT-PCR and normalized with TBP mRNA. Data are plotted relative to the wild-type cell line D422. (D–F) APRT enzymatic activity was determined for mutants S23 (D), S62 (E), and S1 (F) and their repaired counterparts obtained after transfection with the corresponding repair-PPRHs and subsequent selection. In (F), also shown are the APRT enzymatic activity levels of three clones repaired with the long-distance repair-PPRH (LD-HpS1E2rep). The D422 wild-type cell line was included in the determination as the positive control. Data are represented as mU of APRT enzyme divided by the mg of total protein extract. Error bars represent the standard error of the mean of three experiments. Statistical analysis was performed comparing the mean value of each clone with the mean value of the mutant sample. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.