genomewide characterization of host-pathogen interactions by ...

Maren Depke
Material and Methods
Pathogen Gene Expression Profiling
In vitro Infection Experiment Study
Host cell line and culture conditions [performed by Petra Hildebrandt]
S9 cells were cultivated as described above (see Material and Methods/Host Cell Gene
Expression Pattern in an in vitro Infection Model, page 51).
Bacterial growth medium and cultivation [performed by Melanie Gutjahr and Maren Depke]
Staphylococcus aureus RN1HG or RN1HG GFP (plasmid pMV158GFP) was grown in 200 ml
pMEM medium at 37°C with linear shaking of 110 strokes/min (stroke length 28 mm) in 1000-ml-
Erlenmeyer bacterial culture flasks. All other experimental parameters (e. g. medium pMEM)
were identical to those described above (see Material and Methods/Host Cell Gene Expression
Pattern in an in vitro Infection Model, page 51).
Cell culture infection model [performed by Melanie Gutjahr and Maren Depke]
The cell culture infection model was performed as described above (see Material and
Methods/Host Cell Gene Expression Pattern in an in vitro Infection Model, page 51) and also
included the same two time points of 2.5 h and 6.5 h after start of infection (Fig. M.5.2).
cultivation to OD 0.4
experimental time point / h
0 1 2 3 4 5
6 7
infection
1h
lysostaphin treatment
t 0 1 h
2.5 h 6.5 h
internalized
bacterial
samples
internalized
S. aureus
internalized
S. aureus
Fig. M.5.2:
Time line
of infection
experiments
for bacterial
samples.
bacterial
control
samples
exponential
growth
phase
(OD 0.4)
S. aureus
nonadherent
S. aureus
serum/CO 2
control
S. aureus
anaerobic
S. aureus
serum/CO 2
control
S. aureus
serum/CO 2
control
S. aureus
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