FIGURE 9.

EzrA is in complex with Z-ring regulators and cell-cycle regulators. Associations of EzrA with FtsZ-ring regulators (MapZ, FtsA, SepF, ZapA) and cell-cycle regulators (DivIVA, FtsK, StkP) in Spn were tested by co-IP in (A) and (B) or B2H in (B). Experiments are representative from two independent biological replicates. (A) Top panels, labeling of transferred membranes with anti-FLAG in the eluted fractions shows the presence of FLAG-tagged bait protein. Middle panels, labeling with anti-HA in the input fraction shows the presence of EzrA-HA. Bottom panels, labeling with anti-HA in the eluted fraction shows if EzrA-HA was associated with the bait protein. Strains were subjected to Co-IP and Western blotting as described in section “Materials and Methods.” Complete genotypes are listed in Supplementary Table 1. (I) “No bait” (IU9713), DivIVA-FLAG³ (IU11414), MapZ-FLAG³ (IU11430), FLAG-FtsA (IU11476), FtsK-FLAG² (IU11664. (II) “No bait” (IU6810), SepF-FLAG (IU12076), or StkP-FLAG² (IU12077). (III) “No bait” (IU11939) and ZapA-FLAG (IU11840). (B) Summary of EzrA co-IP results and pairwise B2H interaction assays. See Tables 2, 3 for quantitation and tabulation of co-IP results. See Supplementary Figure 21 for B2H results.