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. Author manuscript; available in PMC: 2014 Jun 12.
Published in final edited form as: J Cell Sci. 2014 Jan 14;127(0 5):1052–1064. doi: 10.1242/jcs.141226

Figure 7. Antigen uptake by protrusive podosome-like structures.

Figure 7

(A) Schematics of the control experiments for passive leakage of quantum dots or OVA-Alexa fluor 647 (OVA-647) through the filters. (B–C) Leakage assay of quantum dots (B) or OVA-647 (C) through filters with different pore sizes and with or without gelatin impregnation. (D) Scheme of the antigen uptake experiments. (E) Confocal images of dendritic cells cultured on gelatin-coated filters with 1 μm pore sizes. A suspension of quantum dots linked to gp120 (Qdot; left; magenta) or a solution of OVA-647 (right; magenta) was applied to the other side of the filter. The cells were stained with phalloidin-Alexa fluor 488 (Phal; green) and imaged after 1 hr incubation (see Fig. 8A–B for quantification). (F) Live cell imaging of dendritic cells tranfected with LifeAct-GFP and cultured on filter. At time t = 0, OVA-647 was applied to the other side of the filter. The inset shows the increase of OVA-647 fluorescence in time at the position of 3 actin cores marked with orange arrow heads (i – iii). Full dataset is in Supplementary Movie 3. (G) Time course of OVA-647 uptake for dendritic cells on filter treated with 20 μM Pitstop 2 (red) or Pitstop 2-negative control (black) (± SEM of three independent repeats). Scale bars: E, 10 μm; F: 2 μm.