Extended Data Fig. 8: Western immunoblots revealing the interactions between GpsB, PBP1a, PBP2a, S Protein, MacP, MpgA and PgdA.
All Samples were analyzed by immunoblotting using anti-GFP for the detection of bait proteins and either anti-PBP1a, anti-PBP2a, anti-S Protein, anti-MacP, anti-MpgA, anti-PgdA or anti-GpsB antibodies to determine the presence of the corresponding protein. The data shown are representative of three independent experiments. (a) Co-immunoprecipitation of PBP1a, PBP2a, S Protein, MacP, MpgA and PgdA with GFP-GpsB and GFP (control) using anti-GFP antibodies. The green triangle indicates the GFP-GpsB bait. (b) Co-immunoprecipitation of PBP1a, PBP2a, S Protein, MacP, MpgA and PgdA with either GFP-MacP or GFP-S Protein in WT (left panel) or ∆gpsB (right panel) cells. The blue and grey triangle indicate the GFP-MacP and mGFP-S Protein baits, respectively. Native S Protein and MacP are indicated by black triangle whereas their respective GFP-fusions are indicated by green triangle. (c) Co-immunoprecipitation of PBP1a, PBP2a, S Protein, MacP, MpgA and PgdA with either GFP-PgdA or GFP-MpgA in WT (left side) or ∆gpsB (right side) cells. The blue and grey triangle indicate the GFP-PgdA and mGFP-MpgA baits, respectively. Native PgdA and MpgA are indicated by black triangle whereas their respective GFP-fusions are indicated by green triangle. (d) Interaction diagram of GpsB corresponding to (a). Weak interactions are shown as red dashed lines, and strong interactions as red solid lines (e) Interaction diagram of MacP and S Protein corresponding to (b). Weak interactions are shown as black dashed lines, and strong interactions as black solid lines. Interactions strongly affected in absence of GpsB are represented by blue dashed lines. (f) Interaction diagram of PgdA and MpgA corresponding to (c). Weak interactions are shown as black dashed lines, and strong interactions as black solid lines. Interactions strongly affected in absence of GpsB are represented by blue dashed line. (g) Interaction diagram combining data from (d–f).
