Fig. S1. Aerobic growth of a ?lasB mutant and SDS-PAGE analysis of its culture supernatants.

1
Fig. S1. Aerobic growth of a ?lasB mutant and
SDS-PAGE analysis of its culture supernatants.
The mutant bacteria grown overnight in LB at 37
C were inoculated at 1100 in LB 0.4 NO3-,
and growth was monitored by measuring OD600.
Aliquots of the culture were harvested every hour
and protein contents present in each culture
supernatant (CS) were analyzed by SDS-PAGE. For a
comparison, SDS- PAGE of the PAO1 CS harvested
after 8 hr aerobic growth was shown to the right.
The protein band that corresponds to the mature
elastase is shown with an arrow.
2
Fig. S2. The effect of anaerobiosis on the
elastase secretion of various P. aeruginosa
strains including clinical isolates. (A) Strains
indicated at the top were grown in LB plus 0.4
(w/v) NO3- either aerobically (21 O2) or
anaerobically (0 O2) for 18 hrs. The level of
elastase was analyzed by SDS-PAGE. PA14 and PAK
are non-mucoid pathogenic strains, while FRD1 is
a mucoid laboratory strain derived from a CF
patient. Pneu1, 2 and 3 are non-mucoid isolates
from Korean pneumonia patients. (B) MTT cell
viability assay of A549 human epithelial cells
treated with cell-free culture supernatants (CS)
of indicated strains. A549 cells were treated
with the same set of CSs as in panel A.
Experimental conditions are identical to those
described for Fig. 1A. plt0.01 vs. treatment with
anaerobic CS.
3
Gene name Primer sequences (5'-3')
mvfR F ATCAAGCAGGACAACGCGGA R GCAGGGAGGCATTGCACAAC
rsaL F ACAGCCCCAAAACATGGCCT R GGGCAGGTTCTCGCCATTCT
vqsR F AGCAGATCGCGCTGTTCGAG R CTCCTCCCTGACCGCATCCT
rhlR F GCGCTTTCACATCGACCAGG R GCGGTGGTGTATTCGTCCCA
rhlA F GCGCTTTCACATCGACCAGG R GCGGTGGTGTATTCGTCCCA
PA3904 F GGTAGGTAGCGTCGGCACCC R GGCATTGCAGGCACAGCTTC
PA4677 F CGAAGCGGTGTTCACCAAGC R GCCGTAACGCTGCAACAACC
xcpP F GCGCGGACGACATTACAAGC R CGAGTCTTCTTCGGCGGGTT
pqsA F CCACTCCGCTGGACGACAAC R GCAGCATGTGCGAGGGAATC
pqsC F ATCTGTTCGGCTTCCTCGCC R CGCACTCCATCTGCGAATCC
lasB F CCGCAAGACCGAGAATGACA R CTTCCCACTGATCGAGCACT
lasI F TTCAAGGAGCGCAAAGGCTG R GTTCTTCAGCATGTAGGGGC
rhlI F CTTCATCGAGAAGCTGGGCT R AGGTAGGCGAAGACGTCCTT
rpoD F AAGGCCCTGAAGAAGCACGG R GATCGGCATGAACAGCTCGG
Table S1. Primers used for qRT-PCR