Journal: Journal of cell science

Article Title: Podosomes of dendritic cells facilitate antigen sampling

doi: 10.1242/jcs.141226

Figure Lengend Snippet: (A–B) Confocal images of dendritic cells cultured on glass (A) and on filters with 1 μm pore size (B). Actin was labeled with phalloidin-Alexa fluor 546 (Phal; magenta) and clathrin was visualized by immunostaining (AB; green). The filters were impregnated with Alexa fluor 633-labeled gelatin (Filter; grey). The yellow line indicates the position of the orthogonal view. Yellow arrow heads indicate randomly selected actin-rich cores. The red arrow head indicates the approximate filter surface. (C) Quantification of the fraction of clathrin positive actin cores from panels A–B. (D–F) Same as panels A–C, but now with immunostaining for DC-SIGN, DCIR, Dectin-1, CD206 and CD71. Error bars show the spread of data for multiple cells from at least two independent experiments. Scale bars, 2 μm.

Article Snippet: The following primary antibodies were used for the immunofluorescence: mouse anti-vinculin (V9131, Sigma) at 1:200 dilution (v/v), mouse anti-talin (T3287, Sigma) at 1:100 (v/v), mouse anti-paxillin (349 | MAB3060, Sigma) at 1:100 (v/v), mouse anti-CD11b (ITGAM or Bear-1) (IM2581, Coulter) at 1:200 (v/v), mouse anti-CD29 (or ITGB1) (clone ts2/16) at 1:200 (v/v), mouse anti-MMP-14 (MAB3328, Bio Connect / MilliPore) at 1:100 (v/v), rat anti-tubulin (ab6161, Abcam) at 1:500 (v/v), mouse anti-human CD209 (or DC-SIGN) (551186, BD Bioscience) at 1:200 (v/v), mouse anti-human DCIR (DDX0180, Dendritics) at 1:200 (v/v), mouse anti-human Dectin-1 (MAB1859, R&D Systems) at 1:100 (v/v), mouse anti-human CD206 (555953, BD Biosciences) at 1:250 (v/v), mouse anti-human CD71 (347510, SALK) at 1:50 (v/v), mouse anti-clathrin light chain (C1985, Sigma) at 1:100 (v/v) and mouse anti-human MHC class II (clone Q5/13 ( Quaranta et al., 1980 )) at 20 μg ml −1 .

Techniques: Cell Culture, Labeling, Immunostaining