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. 2019 Sep 10;10:2133. doi: 10.3389/fimmu.2019.02133

Figure 2.

Figure 2

CLCF1 activates LSK cells in an indirect fashion. (A) WBM (upper panel) or FACS-sorted LSKs (lower panel) stimulated with IL-6 (50 ng/ml) or CLCF1 (100 ng/ml). Filled gray histograms represent STAT3 phosphorylation in unstimulated cells. (B) The experimental design used to assess the indirect effect of CLCF1 on BM-derived cell. WBM cells were incubated with PBS or CLCF1 for 4 h, washed then cultured for another 24 h. CM was then transferred to stimulate freshly isolated BM cells for 24 h prior to LSK quantification by flow-cytometry. (C) Absolute LSK counts from the experiment shown in panel B. CLCF1 or CLCF1-derived CM is represented by the black bars whereas white bars represent the PBS control condition. (D) Cytokine/chemokine analysis of CM collected from cells incubated with PBS or CLCF1 for 24 h. **P < 0.01, ***P < 0.001.