Response of human REV3 gene to gastric cancer inducing carcinogen N-methyl-N’-nitro-N-nitrosoguanidine and its role in mutagenesis

Figure 1 The detection system of untargeted mutation occur-ring on undamaged DNA template. Cells were pretreated by MNNG, and then intact and undamaged shuttle plasmid pZ189 was transfected into cells after removing MNNG. After repli-cation for 48 h in cells, replicated pZ189 plasmid was rescued and then transformed to host bacterial MBM7070 to screen pZ189 mutants.

Figure 2 The response of the human REV3 to MNNG at different time points. The response of REV3 to MNNG was measured in human 293 cells and FL cells at the transcriptional level by using RT-PCR. ^aP < 0.05, compared with the control.

Figure 3 Detection of antisense REV3 RNA fragment expressed in 293-REV3^- cells using RT-PCR. A, 100 bp DNA ladder; B, RT-PCR result in Dex-treated 293-REV3^- cells; C, RT-PCR result in 293-REV3^- cells (Dex-free); D, a positive control.

Figure 4 Western blotting showing the loss of REV3 protein in transgenic FL-REV3^- cells. Western analysis showing the level of expression of REV3 protein in nuclear extracts from REV3 antisense-expressing transfectant FL-REV3^- and its parental strain FL. Ku-70 was used as the loading control.