Figure 5
Figure 5. Torin1-exposed DCs produce increased IL-10, but B7-H1 upregulation does not require autocrine IL-6 or IL-10. (A) BM from IL-10-gfp reporter mice was differentiated in RAPA or Torin1 from day 2 to day 8. DCs were stimulated on day 7 where indicated and interrogated for B7-H1 and IL-10 coexpression by flow cytometry. (B) IL-10-gfp histogram for CD11c-gated DCs. MFI is indicated in the upper right corner. (C) The percentage of B7-H1+IL-10+ DCs from panel A was quantified across multiple experiments. (D) Quantification of IL-10 MFI across multiple experiments normalized to DMSO control DCs. Cell-free supernatants were assessed for IL-12/23p40 (E), IL-6 (F), and IL-1β (G). *, # P < .05 compared with DMSO or DMSO+LPS and RAPA or RAPA+LPS, respectively. DCs were differentiated from IL-6−/− (H,I) or IL-10−/− (J,K) BM cells in the presence of Torin1. B7-H1 expression was analyzed on CD11c-gated DCs by flow cytometry and quantified. *P < .05 compared with WT or WT+LPS. Data are from n = 2 to 3 independent experiments. N.D., not detected.

Torin1-exposed DCs produce increased IL-10, but B7-H1 upregulation does not require autocrine IL-6 or IL-10. (A) BM from IL-10-gfp reporter mice was differentiated in RAPA or Torin1 from day 2 to day 8. DCs were stimulated on day 7 where indicated and interrogated for B7-H1 and IL-10 coexpression by flow cytometry. (B) IL-10-gfp histogram for CD11c-gated DCs. MFI is indicated in the upper right corner. (C) The percentage of B7-H1+IL-10+ DCs from panel A was quantified across multiple experiments. (D) Quantification of IL-10 MFI across multiple experiments normalized to DMSO control DCs. Cell-free supernatants were assessed for IL-12/23p40 (E), IL-6 (F), and IL-1β (G). *, # P < .05 compared with DMSO or DMSO+LPS and RAPA or RAPA+LPS, respectively. DCs were differentiated from IL-6−/− (H,I) or IL-10−/− (J,K) BM cells in the presence of Torin1. B7-H1 expression was analyzed on CD11c-gated DCs by flow cytometry and quantified. *P < .05 compared with WT or WT+LPS. Data are from n = 2 to 3 independent experiments. N.D., not detected.

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