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. 2015 Jun 1;212(6):927–938. doi: 10.1084/jem.20142384

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© 2015 Kim et al.

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Figure 3. — Increased IL-18 production from Wdr1^rd/rd CD11b^intCD115⁺ monocytes. (a) BM from WT or Wdr1^rd/rd mice was cultured in L929 cell-conditioned medium for 4 d. Propidium iodide negative (PI^-ve) and CD11b^int/low populations were gated (blue box; left). FSC-A, forward scatter A. Based on CD115(M-CSFR) and Gr1 immunostaining, four subsets of CD11b^int/low populations were sorted by flow cytometry (blue boxes; right). (b and c) 5 × 10⁴ cells from each sorted population were incubated with 1 µg/ml LPS. 48 h after stimulation, culture supernatants were harvested and assayed by ELISA for IL-18 (b) and TNF (c). (d) Sorted populations were cytocentrifuged and examined by microscopy after May-Grünwald Giemsa stain. Bar, 10 µm. All data are representative of two independent experiments; error bars represent SEM of two technical replicates of two biological duplicates. **, P < 0.01; ***, P < 0.001 by unpaired t test.