Fig. 5: Brush localization of translation factor GFP fusions.

a Top: Schematic of the functionality assay for translation proteins fused to GFP with the reporter protein mScarlet. Bottom: GFP and mScarlet FI(t), showing expression of a functional EF-G with a C-terminal GFP fusion versus a non-functional control (dummy protein fused to GFP) and the corresponding reporter expression (replicates see Supplementary Fig. 23). b Scheme: translation proteins were fused to GFP and expressed from surface-immobilized DNA brushes. TIRF microscopy, illuminates a ~100 nm region above the surface, was used to visualize protein localization at the DNA brushes (height ~100 nm). c Mean GFP fluorescence signal averaged over 20 min from TIRF microscopy of DNA brushes encoding various translation protein–GFP fusions. Below: average time-resolved fluorescence traces from three DNA brushes within each cluster. Mean ± s.e.m. (n = 3 DNA brush cluster). “Brush” denotes mean GFP FI within the region of interest (ROI) of the DNA brush; “background” represents mean FI from a neighboring ROI lacking DNA; “epi” corresponds to epifluorescence, which detects GFP throughout the bulk volume above the surface. A second data set in Supplementary Fig. 25. Scale bars = 200 µm. Source data are provided as a Source Data file.