Pr

1
Tran et al, supplemental material
P-cou 2 mM
O-cou 2 mM
pTR
pTC
pTZ
BS
MgCl2
2.5
2.5
0
1
0,25
0.5
0
0,25
0.5
P
PadR
probe
OD600 1,6
Fig. S1. Western-blot of purified polyclonal PadR
antibodies on crude protein extracts from
recombinant E. coli strains. Antibodies were
produced and purified by Eurogentec, Belgium,
from rabbits immunized with PadR purified from
recombinant E. coli strain BL21pER. pTZ, control
strain with the no-insert pTZ19R vector pTR,
pTZ19R harboring and expressing the padR gene
pTC, pTZ19 expressing padC and BS, crude extract
from the B. subtilis wild-type strain (see Table
1 and Fig. 4). Crude extracts were resolved by
SDS-PAGE and proteins were electro-transferred
onto a nitrocellulose membrane prior to
Western-blotting.
Fig. S2. EMSA with the padC promoter probe and
1nM of PadR at different concentrations of MgCl2
from 0 to 2.5mM in the presence of 2 mM of
p-coumaric (P-cou) or o-coumaric (O-cou) acid. P,
padC promoter probe without protein. Effective
inhibition of p-coumaric acid binding was
detectable when MgCl2 concentrations were lower
than 0.25mM. No effect was detected with
o-coumaric acid even in the absence of MgCl2 (For
EMSA conditions, refer to the legend of Fig. 6).