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Blood, Vol. 107, Issue 4, 1537-1545, February 15, 2006
Nitric oxide and cGMP protein kinase (cGK) regulate dendritic-cell migration toward the lymph-node–directing chemokine CCL19
Blood Giordano et al.
107: 1537
Supplemental materials for: Giordano et al
Files in this Data Supplement:
- Table S1. Real-Time PCR Primers (PDF, 15.4 KB)
- Figure S1. Nitric oxide reduces the LPS-induced release of IL-10 and TNFα (JPG, 30.2 KB)
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iDCs were cultured for 24hrs in medium (C), LPS only (0.3µg/ml), or LPS plus 100µM NOR4. Supernatants were collected and analyzed for IL-6, IL-10 and TNF by ELISA. Data represent the mean values ± SEM from different experiments: N=7 for IL-6 and N=4 for Il-10 and TNF. P= 0.014, **P< 0.01, LPS-treated DCs vs. LPS plus NOR4-treated DCs, paired Student’s t-test.
- Figure S2. LPS treatment of induces an increase in cGKI expression in DCs, is enhanced by NOR4 (JPG, 72.8 KB)
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iDCs were stimulated or not (white bars) for 24hrs with graded doses of NOR4 (30-100µM) (grey bars) or LPS (0.3µg/ml) in presence (datched bars) or absence (black bars) of graded doses of NOR4 (30-100µM). mRNA levels of cGKI, cGKII, cGKIa, cGKIb were determined by quantitative real-time RT-PCR. The relative expression of cGK was normalized to the endogenous beta-actin. Data are from one representative experiment out of two giving the same results.
- Figure S3. NOR4 increases cGMP levels in DCs (JPG, 55.1 KB)
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DCs were treated over 24hrs with medium only (o), 100µM NOR4 (r) or 0.3µg/ml LPS only (n) or both LPS and NOR4 (p). At the indicated time points (30’, 1.5h, 5h, 24h) cGMP levels in cell extracts were measured using a cGMP-EIA assay. Data shown are a representative of 2 independent experiments.
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