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Blood, Vol. 111, Issue 4, 1781-1788, February 15, 2008
Bmx tyrosine kinase regulates TLR4-induced IL-6 production in human macrophages independently of p38 MAPK and NF B activity
Blood Palmer et al.
111: 1781
Supplemental materials for: Palmer et al
Files in this Data Supplement:
- Figure S1. Bmx siRNA induces knockdown of Bmx mRNA but not protein (JPG, 49.9 KB)
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Peripheral blood monocytes were transfected with increasing doses (50, 100 and 200nM) of targeting and control siRNA oligonucleotides and differentiated in the presence of 100ng/ml M-CSF in culture for 4 days. (A) Bmx mRNA levels were assessed by real-time PCR. Mean values (±SEM) for 3 separate donors are shown normalised to buffer control (100%). (B) The expression of Bmx and Lamin positive control (Santa Cruz antibody) were assessed by Western blotting.
- Figure S2. Bmx kinase dead and kinase deleted mutants do not act as dominant negatives (JPG, 72.7 KB)
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Macrophages were infected with AdBmx, AdBmx KD (K445E), AdBmx KDel ( 417-675) or Ad0 as before. Cells were cultured in complete medium and stimulated with LPS (10ng/ml), for 18 hours. LPS-induced TNF and IL-6 production was assessed by ELISA. Mean values (±SEM) for 3 separate donors normalised to LPS-treated controls are shown. Statistical significance was assessed using Student’s t-test.
- Figure S3. Bmx does not signal upstream of NF-κB (JPG, 44 KB)
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Mean densitometry units (from Figure 4) pooled for 3 separate donors (±SEM) are shown normalised to 30 min LPS for uninfected controls.
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