Fig. S3.

CdiA-CT^EC536 anticodon nuclease activity is metal-dependent, and the CdiA-CT^EC536 variants interact stably with CysK. (A) Total E. coli RNA was incubated with purified CdiA-CT^EC536 and CysK. Reactions were supplemented with 1 mM MgCl₂ or CaCl₂ where indicated. Reactions were analyzed by denaturing PAGE and ethidium bromide staining. (B) Purified CdiA-CT^EC536 toxins were mixed with CysK-His₆ (input lanes) and subjected to Ni²⁺-affinity chromatography. Proteins that did not bind the Ni²⁺-NTA agarose matrix are shown in free lanes, and the bound lanes contain proteins eluted with imidazole.