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Blood, Vol. 107, Issue 4, 1459-1467, February 15, 2006
Dendritic-cell-associated C-type lectin 2 (DCAL-2) alters dendritic-cell maturation and cytokine production
Blood Chen et al.
107: 1459
Supplemental materials for: Chen et al
Files in this Data Supplement:
- Figure S1. Mouse DCAL-2 gene structure, sequence and protein alignment with human DCAL-2 (JPG, 199 KB)
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(A) The nucleotide and protein sequence of mDCAL-2. The same features are highlighted as in Figure 2. Briefly, the ITIM is outlined and indicated in bold and the putative transmembrane region is underlined. The six cysteine residues that create three disulphide bonds in the CRD are circled and shaded and four potential glycosylation sites are boxed. Only one potential glycosylation site is located within the CRD. (B) The protein alignment of mDCAL-2 with hDCAL-2. The shaded areas show homology between mDCAL-2 and hDCAL-2. At the amino acid level mDCAL-2 has 57% homology with the hDCAL-2.
- Figure S2. hDCAL-2 mRNA is restricted in hematopoietic and myeloid cell lines (JPG, 131 KB)
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(A) Tissue specific expression of hDCAL-2 mRNA. Northern blot analysis of hDCAL-2 expression revealed the highest level of expression in peripheral blood monoleukocytes (PBMCs) and lung. All other tissue, placenta, small intestine, liver, kidney, spleen, thymus, colon, skeletal muscle, heart and brain were negative.  -actin labeling was present for all tissues tested. (B) RT-PCR analysis of human cell lines shows hDCAL-2 is expressed weakly in the HL-60, U937 and Nalm-6 cell lines. Results are representative of three independent experiments.
- Figure S3. DCAL-2 signal did not affect LPS- or zymosan-induced IL-6 or IL-10 expression but selectively inhibit LPS-induced TNF-α expression (JPG, 71.5 KB)
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iDCs were stimulated with or without indicated doses of LPS or zymosan in the presence or absence of anti-DCAL-2 (10 µg/ml) or IgM control. After 24 h stimulation, culture supernatants were harvested and analyzed for the expression of TNF- (A), IL-6 and IL-10 (B) by ELISA. Results were representative of four independent experiments. Mean of triplicate wells ± SEM. *P<0.05; **P<0.01, paired student’s t-test.
- Figure S4. DCAL-2 signal enhanced TNF-α and IL-6 expression by CD40L-matured DCs (JPG, 53.9 KB)
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iDCs were stimulated with CD40L fusion protein (3 µg/ml) in the presence of anti-DCAL-2 or IgM control for 24 h. Supernatants were collected and analyzed for TNF- and IL-6 expression by ELISA. Mean of triplicate wells ± SEM; data are representative of three independent experiments using different donors. Mean of triplicate wells ± SEM. *P<0.05; **P<0.01, paired student’s t-test.
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