Figure 3.

Dendritic cell‐specific intercellular adhesion molecule 3‐grabbing non‐integrin 1 (DC‐SIGN) regulates p65 activation via the myeloid differentiation primary response 88 (MyD88)‐independent pathway. Negative control (NC) or DC‐SIGN siRNAs were transfected into human kidney 2 (HK‐2) cells, and treated or untreated with lipopolysaccharide (LPS) (100 ng/ml) for 60 min. (a) Western blotting was used to detect the DC‐SIGN knock‐down efficiency and p65 phosphorylation, which was quantified by densitometry in three independent experiments as relative units (DC‐SIGN/α‐tubulin, p65 phosphorylated protein/total protein). (b) The immunofluorescence assay used to detect p65 nuclear translocation. (c) Western blotting was used to detect IKB kinase (IKKɛ), TANK binding kinase 1 (TBK1), p38 and c‐Jun N‐terminal kinases (JNK) phosphorylation with DC‐SIGN knock‐down, which was quantified by densitometry in three independent experiments as relative units (IKKɛ, TBK1, p38 and JNK phosphorylated protein/total protein). Data are expressed as the mean ± standard deviation (s.d.) from three independent tests. *P < 0·05; **P < 0·01 compared with LPS untreated; ##P < 0·01 compared with NC in the same group. [Colour figure can be viewed at wileyonlinelibrary.com]