Supplementary Figure 3: Apoptotic cell-driven AhR induction is independent of IDO and driven by TLR9. | Nature Immunology

Supplementary Figure 3: Apoptotic cell-driven AhR induction is independent of IDO and driven by TLR9.

From: Apoptotic cell–induced AhR activity is required for immunological tolerance and suppression of systemic lupus erythematosus in mice and humans

Supplementary Figure 3

(a) mRNA for Ido1 was measured by sqPCR in BMDM and Ap-BMDM. (b) Culture supernatants from BMDM cultures treated as described in a were assessed for IDO enzymatic activity (i.e. increased tryptophan conversion to kynurenine) by HPLC. (c) Cyp1a1 message was measured in BMDM and Ap-BMDM cultures by sqPCR. Groups of Ap-BMDM included B6.Ido1−/− or B6 +/- the IDO-inhibitor D-1-methyl-tryptophan (200μm). (d) Ap-BMDM cultures were done in the presence of oligonucleotide inhibitors of TLR7 (IRS 661), TLR9 (IRS 869), or TLR7/9 (IRS 954) (1μM for all inhibitors). Cyp1a1 expression was measured by sqPCR. For a,b,d,d n=3 biologically independent samples per group. Bars and data points in line graphs are mean values for group +/- standard deviation. *=P≤ 0.05, ns= not significant as determined by two-sided Student’s t-test. For sqPCR samples were normalized against Bactin. (e) Full image for immuno-blots shown in Fig. 1c. (f) Full image for immuno-blots shown in Fig. 3b. (g) Full image for immuno-blots shown in Fig. 3d. All Experiments were repeated three times with similar results.

Back to article page