Fig. 2

Transgelin-2 is essential for global actin rearrangements in activating DCs. a SEM images of WT or Tagln2^−/− DCs in the resting state. b Localization of transgelin-2 in DCs. Enlarged boxed images show the localization of transgelin-2 in the filopodial tip at the edge of a protrusive region (left) and podosomes, along with F-actin and vinculin (right). The fluorescence intensity profiles of each protein were analyzed using Fluoview. Scale bar, 5 μm. c Re-assembly of podosomes in WT or Tagln2^−/− DCs in response to LPS. Podosome cores were visualized by phalloidin staining, and the number of podosomes per cell was counted as described in Materials and Methods. Scale bar, 5 μm. d Effects of transgelin-2-deficiency on DC spreading and actin polymerization. WT or Tagln2^−/− DCs were seeded on Fn-coated coverslips and stained with phalloidin-TRITC. Cell spreading areas were calculated using ImageJ software. F-actin contents were analyzed by flow cytometry. Data from c and d represent the mean of three experiments ± SEM. *P < 0.05, **P < 0.001. e Determination of signaling cascade in WT or Tagln2^−/− DCs in response to CCL19 (200 ng/mL) by western blot. Results are representative of three independent experiments