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. Author manuscript; available in PMC: 2022 Nov 30.
Published in final edited form as: Nat Microbiol. 2022 May 30;7(6):856–867. doi: 10.1038/s41564-022-01132-w

Fig. 3. Biochemical characterization of the interaction between Nocardioides Leaf374 and Aeromicrobium Leaf245.

Fig. 3

a) Nocardioides Leaf374 colonization in mono-association or in combination with Leaf245. Strains were sequentially inoculated and harvested as indicated below. Leaf374 CFU were enumerated on MM maltose agar plates. Shown are the median and individual data points of log10-transformed CFU per gram fresh weight (FW) (n= 12). P-values (two-sided Wilcoxon rank sum test) and log2 fold-changes compared to mono-association are indicated. For colonization levels of Aeromicrobium Leaf245 see Supplementary Fig. 6. d: days after planting b) In vitro interaction between Leaf374 (in overlay agar) and Leaf245 live cells and cell free supernatant (spotted on top). To increase the contrast of the inhibition area, plates were imaged on top of a checkerboard background. c) Fractionation of Leaf245 supernatant by size exclusion chromatography. Shown is the absorption trace at 215 nm. Top right box shows trace of the complete run and marks the section that is enlarged in the main panel. Solid red lines indicate the collected fractions and the corresponding bioactivity is shown below. ++: complete-, +: partial- and -: no clearance of Leaf374 overlay agar. Dotted red lines indicate the fractions selected for SDS-PAGE. d) SDS-PAGE protein bands observed in the indicated fractions of Leaf245 supernatant (see panel c) and a second biological replicate. Arrows indicate protein bands that were identified by mass-spectrometry. e) In vitro activity of purified candidate proteins against Leaf374 in overlay agar. Proteins that could not be expressed in sufficient amounts are not shown.