Fig 2: IL-6 production is enhanced in MCPIP3-deficient pDCs.a IFN-a secretion by Zc3h12c+/+(blue)/Zc3h12c-/- (red) plasmacytoid dendritic cells (pDCs) was measured by ELISA after stimulation (n = 3 mice per group). b Il6 expression in Zc3h12c+/+/Zc3h12c-/- FL3TL-pDCs was measured by qPCR (n = 4 mice per group). c Il6 expression in sorted Zc3h12c fl/fl/Zc3h12cDC? bone marrow (BM) pDCs was measured by qPCR (n = 4 mice per group). d IL-6 secretion by Zc3h12c+/+/Zc3h12c-/- FL3TL-pDCs was measured by ELISA (n = 4 mice per group). e IL-6 secretion by sorted Zc3h12cfl/fl/Zc3h12cDC? BM pDCs was measured by ELISA (n = 4 mice per group). f IL-6 secretion by Zc3h12cfl/fl/Zc3h12cDC? splenic cDCs were measured by intracellular staining after R848 activation (n = 4 mice per group). g TNF production by Zc3h12c+/+/Zc3h12c-/- BM pDCs was measured by intracellular staining after 4 h of stimulation (n = 4 mice per group). h TNF secretion by sorted Zc3h12cfl/fl/ Zc3h12cDC? BM pDCs was measured ELISA (n = 4 mice per group). i Il12p40 expression in Zc3h12c+/+/Zc3h12c-/- FL3TL-pDCs was measured by qPCR (n = 4 mice per group). Data are representative of six (a), three (b–e), or two (f–i) independent experiments (mean ± S.D.; two-tailed Student’s t test). Source data are provided as a Source Data file.

Fig 3: Exposure to pDC-derived IL-6 suppresses macrophage TNF and IL-12p40.a Sorted WT bone marrow-derived macrophages (BMDMs) and FL3TL-pDCs were placed into the same wells for 16 h before addition of R848. TNF production in macrophages was measured by intracellular staining in macrophages at 4 h (n = 4 mice per group). b WT BMDMs were treated with recombinant IL-6 at various concentrations, and macrophage TNF or IL-12p40 production was measured by intracellular staining at 4 h. Percentage changes was normalized against WT samples without IL-6 (n = 3 mice per group). c, d Sorted WT FL3TL-pDCs were treated with R848 for 24 h, and their supernatants (pDCsup) or rIL-6 (50 ng/ml) were added to Zc3h12c+/+(blue)/Zc3h12c-/- (red)BMDMs along with R848. TNF production in macrophages at 4 h (c) or 16 h (d) was measured by intracellular staining. Percentage changes was normalized against Zc3h12c+/+ samples without pDCsup or rIL-6 (n = 4 mice per group). e TNF production by Zc3h12c+/+/Zc3h12c-/- BMDMs with sorted WT BM pDC supernatants was measured by intracellular staining after R848-treatment at 4 h (n = 5 mice per group). f TNF production by Zc3h12c+/+/Zc3h12c-/- BMDMs with titrated concentrations of R848-treated sorted Zc3h12c-/- FL3TL-pDC supernatants, with or without IL-6 neutralizing antibodies was measured by intracellular staining at 4 h (n = 3 mice for Zc3h12c+/+; n = 4 mice for Zc3h12c-/-). g IL-12p40 production by Zc3h12c+/+/Zc3h12c-/- BMDMs with pDCsup or rIL-6 was measured by intracellular staining at 4 h (n = 5 mice for Zc3h12c+/+; n = 4 mice for Zc3h12c-/-). h IL-12p40 production by Zc3h12c+/+/Zc3h12c-/- BMDMs with titrated concentrations of R848-treated sorted Zc3h12c-/- FL3TL-pDC supernatants, along with or without IL-6 neutralizing antibodies was measured by intracellular staining at 4 h (n = 4 mice per group). Data are representative of five (c, d), three (g), or two (a, b, e, f, h) independent experiments (mean ± S.D.; two-tailed Student’s t test). Source data are provided as a Source Data file.

Fig 4: Early TNF production is impaired in MCPIP3-deficient macrophages.a Tnf expression in R848-treated Zc3h12c+/+(blue)/Zc3h12c-/- (red) bone marrow-derived macrophages (BMDMs) was measured by qPCR at indicated timepoints (n = 3 mice per group). b, c TNF(b) and IL-6(c) secretion by R848-treated Zc3h12c+/+/Zc3h12c-/- BMDMs was measured by ELISA at indicated timepoints (n = 3 mice for Zc3h12c+/+; n = 4 mice for Zc3h12c-/-). d–f TNF production by Zc3h12c+/+/Zc3h12c-/- BMDMs (d, n = 7 mice for Zc3h12c+/+; n = 8 mice for Zc3h12c-/-), Zc3h12cfl/fl/Zc3h12cMF? BMDMs (e, n = 3 mice for Zc3h12cfl/fl; n = 5 mice for Zc3h12cMF?), and purified Zc3h12c+/+/Zc3h12c-/- or Zc3h12cfl/fl/Zc3h12cMF? peritoneal macrophages (Per.MF) (f, n = 3 mice for Zc3h12cfl/fl; n = 5 mice for Zc3h12cMF?) was measured by intracellular staining after 4 h of stimulation with indicated ligands. g Il12p40 expression in R848-treated Zc3h12c+/+/Zc3h12c-/- BMDMs was measured by qPCR at various timepoints (n = 3 mice per group). h, i IL-12p40 production by Zc3h12c+/+/Zc3h12c-/- BMDMs was measured by intracellular staining after 4 h (h, n = 4 mice per group) or 16 h (I, n = 4 mice for Zc3h12c+/+; n = 3 mice for Zc3h12c-/-) stimulation with indicated ligands. Data are representative of eight (d, e), four (a, b), three (h, i), two (c, f, g) independent experiments (mean ± S.D.; two-tailed Student’s t test). Source data are provided as a Source Data file.