Figure 6

Tsc1 maintains sufficient numbers of antigen-pulsed DCs in antigen-specific T-cell responses. (a) Purified DCs were administered Eα antigen for 6 h, and the efficiency of the formation of peptide–MHC class II complexes was determined by staining with the Y-Ae antibody (n=3). (b) Autophagy detection of splenic DCs, treated with or without RAPA (50 ng/ml) for 16 h. (c) Expression of CCR3, CCR5, CCR6 and CCR7 on splenic DCs. (d) Annexin V and PI staining of splenic DCs (n=3). (e) Ki67 staining of splenic DCs (n=3). (f) Apoptosis of splenic DCs in DC-OT-II co-culture system, after being co-cultured for 3 days(n=5). (g) The percentage of FITC⁺ cells in draining LNs. The skin of the scapular region of WT and CD11c^CreTsc1^f/f mice was painted with 20 μl of 1% FITC. After 48 h, draining LNs were collected and analyzed. (h) Numbers of CFSE⁺ cells in the spleen and LNs from recipients B6.SJL (CD45.1), 3 days after being transferred with CFSE-stained 1 × 10⁶ WT or Tsc1-deficient DCs by i.v. (n=5). *P<0.05, **P<0.01; error bars represent S.D.; all data are representative of at least three independent experiments