Supplemental figures and table for additional data and strain information.

Fig A. Sequence alignment of VgrG proteins in A. dhakensis SSU. The three VgrG proteins were aligned using the BLAST Multiple Alignment tool. Alignment was downloaded in Clustal format and visualized using ESPript with default settings (https://espript.ibcp.fr). The predicted structure of VgrG1, generated by Phyre2, was used as structural template in ESPript. Fig B. Pull-down analyses of A. dhakensis effector-structural protein interactions. a, Pull-down analysis of structural proteins with the catalytically inactive TseI^HFH-AAA. The full-length protein and the cleaved C-terminus of TseI are indicated. b, Pull-down analysis of structural proteins with the effector TseC. c, Pull-down analysis of TssA with TseI^HFH-AAA. d, Pull-down analysis of TssA with TseC. For all pull-down assays, effectors carry a C-terminal 3V5 tag and bait proteins are fused with an N-terminal 6His tag. His-sfGFP serves as a negative control for nonspecific interaction. All proteins were individually expressed in E. coli and cell lysates mixed in pairs. Samples were detected by Western blotting analysis using the anti-V5 and anti-His antibodies, respectively. All pull-down experiments were performed at least twice and a representative result is shown. Fig C. Pull-down analyses of V. cholerae effector-structural protein interactions. a, Pull-down analysis of V. cholerae structural proteins with the catalytically inactive TseL^D425A. b, Pull-down analysis of V. cholerae structural proteins with the effector VasX. c, Pull-down analysis of V. cholerae TssA with TseL^D425A. d, Pull-down analysis of V. cholerae TssA with VasX. For all pull-down assays, effectors carry a C-terminal 3V5 tag and bait proteins are fused with an N-terminal 6His tag. His-sfGFP serves as a negative control for nonspecific interaction. All proteins were individually expressed in E. coli and cell lysates mixed in pairs. Samples were detected by Western blotting analysis using the anti-V5 and anti-His antibodies, respectively. All pull-down experiments were performed at least twice and a representative result is shown. Table A. Plasmids, strains and primers.

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