Fig. 1: ASOs silence ΦKZ transcripts in Pseudomonas.

a, ASOs are delivered into bacterial cells by a CPP. Upon phage infection, ASOs bind the target phage mRNA at the RBS, preventing translation. If the encoded protein is essential, phage propagation is halted, protecting the bacterium (red shield). b, The chmA locus and translation initiation region. ASOs targeting the RBS and the non-targeting control ASO (Ctrl) are depicted. c, PAO1 cells were pretreated with 6 µM ASOs for 30 min before ΦKZ infection at an MOI of 5. Cells were collected at the indicated time points and ChmA was quantified by immunoblotting. A Coomassie-stained gel served as loading control. One representative example of four independent experiments is shown. d, PAO1 cells were pretreated for 30 min with 6 µM ASOs. After ΦKZ infection at an MOI of 0.0001, cells were incubated for 180 min. The phage–cell suspension was spotted on LB plates to assess the number of CFUs and on LB plates with a PAO1 lawn to assess PFUs. A high phage titre causes lysis of nearby bacterial cells after spotting, leading to a lack of CFUs at low dilutions in the control. A representative result of two independent experiments is shown. e, PAO1 cells were pretreated with 8 µM ASOs for 30 min and infected with ΦKZ at an MOI of 10, followed by chemical crosslinking and staining of membranes (FM4-64) and DNA (DAPI) at either 5 or 30 min p.i. f, PAO1 cells were pretreated with 8 µM ASO targeting the phage spindle gene phuZ or a control ASO for 30 min, infected with an MOI of 10 and imaged as in e. In parallel, PAO1 cells were infected with a phuZ deletion phage at an MOI of 0.001. Quantification in Extended Data Fig. 2a. Uncropped images of blots are presented in Supplementary Fig. 1.