Nature - USA (2020-08-20)

Article

b

10 -3 10 -2 10 -1 100 101 102

0.1

0.4

0.03

rifampicin (uM)

OD

600

K-12

K-12 pBADpbgA

pbgA-pBADpbgA + ind.

pbgA-pBADpbgA - ind.

rifampicin (uM)

OD

600

10 -3 10 -2 10 -1 100 101 102

0.02

0.2

K-12

clsABC

c

150000

100000

50000

0

600 800 1000 1200 1400 1600

K-12

ΔclsABC

CL

PE/PG

1376

m/z

normalized intensit

y

1402

d

e

K-1

2

0

10000

20000

30000

lipid

A AUC/μg protein

K-12

0

200000

400000

600000

800000

CL

AUC/μg protei

n

0

200000

400000

600000

800000

CL

AUC/μg protein

K-12

0.52 40 .5 24 0.52 40 .5 24 hours

100

102

104

106

108

liver spleen

total CFU recovere

d

UPEC UPEC

a

-4 -1

cyclophosphamide 0.5 hpi 24 hpi

Extended Data Fig. 1 | In vivo and in vitro characterization of E. coli ΔpbgA
and ΔclsABC strains. a, CFUs recovered from UPEC and UPEC ΔpbgA in
neutropenic mouse tissues after intravenous injection of BALB/C mice 0.5 and
24 h after injection (n = 5 per group). Data are mean ± s.d. with dashed line
indicating lower boundary of detection. b, Rifampicin sensitivity assay with
conditional E. coli K-12 ΔpbgA::pBADpbgA strain. Data are mean ± s.d. for at each
rifampicin concentration for n = 3 of each strain. c, Rifampicin sensitivity assay
with E. coli K-12 and ΔclsABC strains. Data are mean ± s.d. for each rifampicin

concentration for n = 3 of each strain. d, Quantification of lipid A and

cardiolipin measured by MALDI–TOF and Qtrap liquid chromatography–

tandem mass spectrometry (LC–MS/MS), respectively, normalized to total

protein amounts in whole cells (left and middle) or outer membrane vesicles

(right). AUC, area under the curve. Data are mean ± s.d. for each strain for n = 3

replicates. e, MALDI–TOF mass spectrometry analyses detected no cardiolipin

in the ΔclsABC strain (orange) compared to the E. coli K-12 strain (black) when

analysed under matched conditions. Representative results are shown.