1. Table E1 – Sequences of primers used in qPCR experiments
Forward
Reverse
Human
Mineralocorticoid receptor (NR3C2)
GAGGCTTCAGGATGCCATTA
TCTGCAAGCAGGACAATTCTT
Glucocorticoid receptor (NR3C1)
AGAAAACTCCAGCCAGAACTG
GATTTCAGCTAACATCTCGGGGA
18S
GGACACGGACAGGATTGACA
ACCCACGGAATCCAGAAAGA
Alpha 1 D adrenoceptor
TCTGCTGGTTCCCTTTCTTC
CACGCAGCTGTTGAAGTAGC
Mouse
Mineralocortcoid receptor (NR3C2)
CCAGAAGAGGGGACCACATA
GGAATTGTCGTAGCCTGCAT
TTCGCAGGCCGCTCAGTGTT
TTGGGAGGTGGTCCCGTTGCT
α1-a adrenoceptor
GCGGTGGACGTCTTATGCT
TCACACCAATGTATCGGTCGA
α1-b adrenoceptor
CCT GGTCATGTACTGCCG
GACTCCCGCCTCCAGATTC
α1-d adrenoceptor
AGTTGGTGACCGTCTGCAAGT
CGCTGTGGTGGGAACCGGCAG
Endothelin 1 receptor A
CCCAAACCTCCCAAGTCTCTC
TGGAAATGACATGGGCGGTAT
Angiotensin II Receptor type 1
GACTGGATGATGCTGTCTGGC
CTTGGAGGGTTGCTGTGAGT
CGCCGCTAGAGGTGAAATTC
TCTTGGCAAATGCTTTCGC
Table E1 – Sequences of primers used in qPCR experiments

2. Control
LPS
LPS-Aldosterone p
SBP
EC50 (mg/kg)
0.020±10-3
0.019±10-3
0.022±1.2x10-3 ns
Emax (mmHg)
38.15±2.1
31.03±2.5
33.23±2.6
DBP
0.016±10-3
0.009±10-3
0.013±10-3
38.97±1.9
28.63±2.7
30.29±2
0.037*
Table E2 – Effect of Aldosterone on Blood Pressure response to Phenylephrine in Endotoxic Shock. Systolic and Diastolic response to increasing doses of phenylephrine. Results are expressed as mean±SD
*Multiple comparison test: p ≤ 0.05 in all cases

3. Control
LPS
LPS-Aldosterone p
EC50 (Log M)
±5 10-9
±6 10-9
±5 10-9
0.03#
Emax 1
1±0.2
0.55±0.1
0.78±0.3
0.002##
Table E3 – Effect of Aldosterone on ex vivo first-order mesenteric artery contractile response to phenylephrine in endotoxemic mice
Dose-response curve parameters. Results are expressed as mean±SD. Vessels were studied 18 hours after LPS (15 mg/kg) and Aldosterone (A mg/kg) injection. n =
1 Normalized on control
#Multiple comparison test: p<0.05 for LPS vs Control, and LPS-Aldosterone vs LPS
## Multiple comparison test p<0.05 in all cases

4. Table E4 – Brief patient and control subject characteristics
Age
Gender
SAPSII
Medical history
Medications
Site of infection
Microorganism
Outcome
Patient 1 77 M 56
Rectal adenocarcinoma
None
Soft tissue
Escherischia coli
Discharged
Patient 2 69 42
Aortic valve replacement
warfarin
Abdomen (pancreatitis) NA
Dead in ICU
Patient 3 55 50
Myasthenia gravis, thymoma
Prednisone, azathioprine, neostigmine
Lung
Patient 4 44 F 35
Seizures
levetiracetam
Patient 5 45
Abdomen
E. coli and E. faecalis
Patient 6 58
COPD
Salbutamol, ipratropium
Str. Peunomniae
Patient 7 32
Chronic sinusitis, ethanol abuse
CNS (meningitis)
Str. Pneumoniae
Control 1 66 -
Control 2
Control 3
Control 4 40
Control 5 38
Asthma in infancy
Control 6 29
Appendicectomy
Levonorgestrel
Control 7 59
Table E4 – Brief patient and control subject characteristics

5. Figure E1 – Effect of Aldosterone on vasoconstrictor receptor expression in endotoxemic mouse aortas. MR and GR mRNA expression in mouse aortas 18 hours after LPS or LPS-Aldosterone injection. Mice were injected intraperitoneally with LPS (15 mg/kg) and Aldosterone (1mg/kg) (n=5 mice per group, * p ≤ 0.05 vs Control, #p ≤vs LPS)

6. Figure E2 – MR and GR expression in endotoxemic mice aortas
Figure E2 – MR and GR expression in endotoxemic mice aortas. MR and GR mRNA expression in mouse aortas 6 hours after LPS or LPS-Aldosterone injection. Mice were injected intraperitoneally with LPS (15 mg/kg) and Aldosterone (1mg/kg) (n=5 mice per group, * p ≤ 0.05 vs Control, #p ≤vs LPS)

7. Figure E3 – MR expression in HUVECs treated with LPS
Figure E3 – MR expression in HUVECs treated with LPS. MR mRNA expression in HUVECs 5, 10 and 24 hours after incubation with LPS (n=5 replicates per group)

8. *
Figure E4 –MR and GR mRNA expression in HUVECs treated with pro-inflammatory cytokines. Cells were incubated with TNF-α (100 ng/ml), IL1-β (50 ng/ml) and IFN-γ (500UI/ml) during 5 hours. (n=6 replicates per condition,* p≤0.05 vs Control)

9. Figure E5 – Time-course and dose-response of MR expression downregulation by TNF-alpha in HUVECs. Panel A - MR mRNA expression in HUVECs 75 to 450 min after incubation with TNF-α (100 ng/ml). Panel B – MR mRNA expression in HUVECs 5 hours after incubation with TNF-α (0.01 to 100 ng/ml) (n=5 replicates per condition, * p≤0.05 vs control) *

10. ns
Figure E6 - GR mRNA expression in HUVECs treated with TNF-α and BAY GR mRNA expression five hours after treatment with TNF-alpha (10 ng/ml) . (n=6 replicates in all groups)

11. Figure E7 – Effect of NF-kappa B inhibitor BAY on MR mRNA expression down regulation by plasma from septic patients Cells were pre-incubated with BAY (10 µM) and incubated with septic/control plasma for 5 hours (n= 3 subjects per group, 3 replicates per condition, *p≤0.05 vs Control, #p≤0.05 vs patients)