Extended Data Fig. 2: Overexpression of degP is associated with improved growth of carbapenem-resistant bacteria.
(a) mRNA levels of degP compared in wildtype and mutant Keio strains, transcript levels were normalized to the level of rrsE. The dotted line delineates degP levels in wildtype (WT), at a fold change of 1. Data are mean ± s.e.m. (WT, n = 7; ∆waaP, n = 4; ∆gmhA, n = 7; ∆atpD, n = 6; ∆hldE, n = 3; ∆waaF; n = 6; ∆yiiS, n = 2; n is defined as biological replicates and each biological replicate consists of at least two technical replicates). Statistical significance was determined using a Kruskal-Wallis test with Dunn’s test for multiple comparisons, relative to the WT (∆waaP, *P = 0.039; ∆gmhA, ****P < 0.0001; ∆atpD, *P = 0.013; ∆hldE, ****P < 0.0001; ∆waaF, ****P < 0.0001; ∆yiiS, ***P = 0.0005). (b) Protein levels of DegP from whole cell lysates in wildtype and mutant Keio strains. RNA Polymerase subunit α served as a loading control. Data is a representative blot of three biological replicates. (c) mRNA levels of degP compared in K. pneumoniae carrying the empty vector (Empty; n = 3) and expressing VIM-2 (VIM-2; n = 3) following growth in vivo. Samples were collected 7 hours post-infection from the blood. Transcript levels were normalized to the level of recA. Data are mean ± s.e.m, and each point represents an individual mouse. Statistical analysis was performed using a two-tailed unpaired t-test, **P < 0.01 (t = 6.727, df=4).
