FIG 2.

Effector-based suppression of SdeA and Ceg6 toxicity. (A) Yeast cells were individually transformed with each member of the L. pneumophila effector repertoire expressed from the pDEST22 plasmid and grown on selective media (data not shown). Each individually transformed cell was grown in liquid culture selecting for pDEST22 plasmid and then pooled. Pools were then transformed with either pDEST32-lpg0208(ceg6) or pDEST32-lpg2157(sdeA). Transformations were plated on SC −Leu, −Trp medium, which selects for both plasmids, and incubated at 30°C. (B) Effectors identified as suppressing Ceg6 or SdeA toxicity through the pooled assay by sequencing were individually transformed into yeast; Rab1A was used as a negative control. Transformed bacteria then underwent a second transformation with Ceg6 or SdeA and were plated on SC −Leu, −Trp medium, which selects for both plasmids. Transformations were incubated at 30°C.