TABLE 2.
Co-immunoprecipitation of complexes containing EzrA-FLAG3 from cross-linked Spn cellsa.
| Prey | Mean ratiob | Detected prey in complex?c | Strains usedd |
| Z-ring regulators | |||
| FtsZ | 22.5 ± 10.9 | Yes | IU6933/IU11602 |
| FtsA | 25.4 ± 9.5 | Yes | IU6933/IU11602 |
| ZapA-HA | 1.2 ± 0.02 | No | IU10267/IU11322 |
| Penicillin-binding proteins (PBPs) | |||
| aPBP2a-HA4 | 1.04 ± 0.12 | No | IU7797/IU11610 |
| aPBP2b-HA | 1.1 ± 0.04 | No | IU6933/IU11602 |
| Cell-cycle regulation | |||
| MreC | 1.0 ± 0.1 | No | IU7797/IU11610 |
aCo-IP experiments were performed as described in section “Materials and Methods.” Prey proteins were detected with anti-HA, anti-FtsZ, Anti-FtsA, or anti-MreC.
bThe mean ratio is determined by dividing Western blot ROI signals of epitope tagged prey in strain expressing ezrA+ by the ROI signal of epitope tagged prey in strain expressing ezrA-FLAG3 ([ROI prey-HA]ezrA+/[ROI prey-HA]ezrA–FLAG3). ROI values are determined as described previously in Materials and Methods. ±, SEM from two independent biological replicates.
cA positive interaction was determined based on the mean ratio value greater than two.
dComplete genotypes are listed in Supplementary Table 1. Strain numbers are listed in the following order; the strain listed first expresses the epitope-tagged prey and untagged EzrA (i.e., IU10302; ftsZ-Myc ezrA+), while the second strain listed contains the epitope tagged prey and EzrA-FLAG3 as the bait (i.e., IU11340; ftsZ-Myc ezrA-FLAG3).