Skip to main content
. 2022 Oct 13;11:e82437. doi: 10.7554/eLife.82437

Figure 4. TseV2 and TseV3 induce DNA double-strand breaks.

Activation of the SOS response was analyzed using E. coli cells harboring the reporter plasmid pSC101-PrecA::GFP and pBRA TseV2 (A) or pBRA TseV3 (B), which were grown in AB defined media with d-glucose or l-arabinose. Data is the mean ± standard deviation (SD) of three independent experiments. ***p < 0.001 (Student’s t-test). Bright-field and DAPI images of E. coli cells carrying pBRA TseV2 (C) or pBRA TseV3 (D) grown in the presence of d-glucose (repressed) or l-arabinose (induced). Results are representative images of three independent experiments. (E, F) Quantification of the mean 4′,6-diamidino-2-phenylindole (DAPI) fluorescence per cell of 200 cells. Data correspond to the mean ± SD of a representative experiment. Scale bar 2 μm. ***p < 0.001 (Student’s t-test). (G) Representative bright-field and GFP images of E. coli coexpressing GamGFP and pEXT20 TseV2 or pEXT20 TseV3. Double-strand breaks appear as foci of GamGFP. Images are representatives of three independent experiments. Scale bar: 5 μm. (H) Quantification of the GamGFP foci shown in (G). Data are shown as the mean ± SD of the three independent experiments. **p < 0.01 (Student’s t-test).

Figure 4—source data 1. Values of GFP signal acquired for the SOS response experiment shown in Figure 4A, B.
Figure 4—source data 2. Values of 4′,6-diamidino-2-phenylindole (DAPI) fluorescence measured for each bacterium, and original images used for quantification shown in Figure 4C–F.
The files can be opened in ImageJ.
Figure 4—source data 3. Original images used to count GamGFP foci shown in Figure 4G, H and numbers of foci.
The image files can be visualized with ImageJ.
Figure 4—source data 4. Original images used to count GamGFP foci in bacteria carrying pEXT20 TseV2 or TseV2D282A in Figure 4G, H.
Figure 4—source data 5. Original images used to count GamGFP foci in bacteria carrying pEXT20 TseV3 or TseV3D230A in Figure 4G, H.
Figure 4—source data 6. Original images used to count GamGFP foci in bacteria carrying empty pEXT20 in Figure 4G, H.

Figure 4.

Figure 4—figure supplement 1. Western blot with anti-GFP antibody of protein extracts from E. coli carrying reporter plasmid pSC101 PrecA::GFP and pBRA with the indicated toxins after induction with 0.2% l-arabinose. anti-DnaK antibody was used as loading control.

Figure 4—figure supplement 1.

Figure 4—figure supplement 1—source data 1. Original images of western blots.