Page 77 - Haematologica March 2020

Cytokine blockade reduces myeloid expansion in RA
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Figure 4. Activation of a proliferation arrest gene program in hematopoietic stem cells from mice with collagen-induced arthritis. (A) Experimental design. (B) Volcano plot showing significantly differentially expressed genes (shaded areas) based on Padj <0.05 (n=3 per group); hematopoietic stem cell (HSC) pools were sorted from three independent sets of mice. (C) Significantly differentially activated upstream regulators as determined by Ingenuity Pathway Analysis. (D, E) Gene ontology (GO) analysis of significantly differentially (D) downregulated and (E) upregulated gene sets. (F-H) Fluidigm gene expression analysis of HSC from control mice (Ctrl) and mice with collagen-induced arthritis showing (F) cell cycle activator genes; (G) cell cycle repression genes; (H) mRNA translation genes. The data are presented as log10 fold expression in CIA HSC versus Ctrl HSC. Ct values were normalized to Actb (n=8-16 per group). The data were compiled from two independent experiments. (I) Experimental design. (J) Representative FACS plots and (K) cell cycle distribution of HSC in Ctrl and CIA mice (n=8 Ctrl and 7 CIA mice). The data were compiled from two independent experiments. See also Online Supplementary Tables S1-S6. *P<0.05; **P<0.01 ***P<0.001, as determined by the Mann-Whitney U-test.
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