Fig. 7: iMoP – a precursor cell between cMoP and monocytes.
From: Monocyte progenitors give rise to multinucleated giant cells
a Volcano plot of Clariom S gene array analysis of cMoP and iMoP. Depicted are −log10(p-value) on the y-axis versus log2(fold change) on the x-axis. Shown are significantly upregulated (red) and downregulated (green) genes in iMoP compared to cMoP. b Heatmaps of gene expression in cMoP and iMoP from Cx3cr1gfp/+ mice. Depicted are standardized log2 differences of significantly up- (red) or downregulated (blue) genes. The analysis was performed by GO-Terms (GO:0045087 innate immune response, GO:0030154 cell differentiation, GO:0007155 cell adhesion, GO:0006629 lipid metabolic process, GO:0016477 cell migration, GO:0019221 cytokine-mediated signaling pathway, GO:0007049 cell cycle). If GO-Terms overlapped, respective genes were only assigned to one group. Affymetrix Clariom S gene array was performed with two biologically independent samples per group (1 = sample 1; 2 = sample 2). c Compilation of over-represented gene-ontology-processes with respect to the significantly upregulated genes in iMoP compared to cMoP. d qRT-PCR analysis of Itgam (Cd11b), Adgre1 (F4/80), Mpo, Tlr13, Msr1 and Kit oncogene mRNA relative to Gapdh mRNA in cMoP, iMoP (red) and MC. Bars show mean ± SEM with n = 3 biologically independent samples. *p = 0.0101 (Tlr13), *p = 0.0142 (Mpo), **p = 0.0038 (Tlr13), **p = 0.0098 (Mpo), ***p = 0.0002 (Itgam, Msr1 cMoP – MC), ***p = 0.0004 (Itgam, Msr1 iMoP – MC), ****p < 0.0001 (ordinary one-way ANOVA, Tukey’s MCT). e cMoP and iMoP (red) were cultured in M3434 Methocult medium. Colonies consisting of at least 10 cells were counted at day 10. Depicted are means ± SEM of n = 4 biologically independent samples examined over two experiments. f qRT-PCR analysis of Ms4a3 mRNA relative to Gapdh in MDP, cMoP, iMoP and MC. Bars show mean ± SEM of n = 4 biologically independent samples examined over at least two experiments. g iMoP isolated from β-actin-gfp+/− mice were transferred intravenously into WT mice. Blood, spleen and bone marrow were analyzed for GFP+ cells (green) 14 h post transfer by FACS for Ly6C, CD11b and F4/80 expression. Representative FACS plots are shown. h Quantification of FACS analysis described in Fig. 7g. Bars show mean ± SEM of n = 5 (blood) and n = 4 (bone marrow, spleen) mice examined over at least three experiments.
