Extended Data Fig. 5: Effect of mutations in Bab and Agp on resistance to T5 and Agp(2-25) induced aggregation of Bab-GFP.
From: Characterization of an amyloid-based antiphage defence system in Escherichia coli
a, Spot assay of E. coli cells of the given genotype infected with 10-fold serial dilutions of T5 in triplicates. Each spot is 4 µL and dilutions are 102 to 108 of the phage stock. Dilutions are from top to bottom and left to right (as depicted in Fig. 2b). b, Western-blot of crude extracts of E. coli MG1655 cells expressing V5-tagged version of Agp and Agp mutants from an arabinose-inducible pBAD vector with an antibody to the V5 tag. As a loading control, the same blot was probed with an antibody to the β’ subunit of E. coli RNA polymerase (αRNAP). c, Western-blot of E. coli MG1655 extracts expressing Bab-GFP or Bab N100P-GFP incubated with various amounts of Agp(2-25) peptide in amyloid conformation and fractionated by centrifugation at 17,000g for 30 min. Agp(2-25) peptide was at 2 mg.mL−1 and was added to 50 µL of crude extract (corresponding to 109 cells). Size of molecular weight markers (lane M) are given in kDa. d, The GFP tag affects antiphage activity of Bab. Spot assay of E. coli cells of the given genotype infected with 10-fold serial dilutions of T5 in triplicates. Each spot is 4 µL and dilutions are 102 to 108 of the phage stock. Dilutions are from top to bottom and left to right (as depicted in Fig. 2b).
