Fig. 7

Recombinant dU-TG2P reconstitutes Tagln2^−/− DC functions. a The transduction efficiency of WT-TG2P (10 µM) in WT or Tagln2^−/− DCs. b Schematic diagram of the domain composition of transgelin-2. A potential ubiquitination site, K78, is highlighted in red. c Stability test of WT-TG2P and dU-TG2P (K78R) in DCs. d, e Reconstitution of transgelin-2 by dU-TG2P. Tagln2^−/− DCs treated with dU-TG2P or heat-inactivated dU-TG2P (H/dU-TG2P) were seeded on Fn-coated plate, and the cell size (d) and spreading (e) were determined by flow cytometry and confocal microscopy, respectively. F-actin was stained by phalloidin-TRITC (E). Cell spreading areas were calculated using ImageJ software. f–h dU-TG2P rescues transgelin-2 function. Tagln2^−/− DCs treated with H/dU-TG2P (10 µM) or dU-TG2P (5–10 µM) were co-incubated with OTII CD4⁺ T cells in the presence or absence of pOVA (323–339), and then, the cells or cultured supernatants were subjected to a conjugates assay (F), a cytokine production assessment (g), and a proliferation test (h). All data from e–h represent the mean of three experiments ± SEM. NS, not significant. *P < 0.01