Figure S1. Effects of cytokine and LTβR blocking on emergency myelopoiesis.

(A) IL-7 receptor expression in proB and preB cells during CFA-induced systemic inflammation. (B) Numbers of developing B-cell subsets in the bone marrow of mice treated with IL-7/aIL-7 for 3 d. (C) Numbers of proB, preB, immature B, and mature B cells in bone marrow of mice injected with CFA (i.p.) and cytokine blocking antibodies over 5 d. (D) Cxcl12-dsRed Geo. mean expression in bone marrow MSCs of mice injected with CFA (i.p.) and cytokine blocking antibodies over 5 d. (E, F) Numbers of proB, preB, immature B, and mature B cells in bone marrow (E) and of neutrophils and inflammatory monocytes in the spleen (F) of mice administered CFA ip and treated with cytokine blocking antibodies. (G) Frequency of BrdU+ cells within the indicated hematopoietic progenitor population, as measured by flow cytometry, of CFA-challenged mice and mice treated with control or LTβR-Ig+ anti-TNF-α blocking reagents. BrdU administration was 1 h before euthanasia. (H) CCL2 protein measurements in bone marrow interstitial fluids recovered from CFA-challenged mice and mice treated with control or LTβR-Ig + anti-TNF-α blocking reagents. Data in all panels are representative of two individual experiments. Bars represent the average, and circles depict individual mice. *P < 0.05; **P < 0.005; and ***P < 0.0005 by an unpaired t test. (I) Principal component analysis distribution plot from RNA sequencing of MSCs in control mice, and in mice challenged with CFA for 2 d treated with Hel-Ig or with LTβR-Ig. (J) GSEA-MSigDB Hallmark pathway analyses from differentially expressed genes. Data are representative of one experiment.