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. 2015 Jun 4;5(1):125–138. doi: 10.1016/j.stemcr.2015.05.008

Figure 4.

Figure 4

Relationship between Hopx Gene Expression and HSPCs

(A) Among 25 HMDP strains, Hopx expression levels in BM MNCs are positively correlated (r = 0.44) with LSKCD150−CD48− frequency. Each data point represents the mean expression of each mouse strain (n = 2–3 mice/strain) across two independent experiments. LSKCD150−CD48−frequency is expressed as a percentage of BM MNCs and expression levels are shown relative to the expression of Hprt, as an endogenous control, in relative units (ru).

(B) Hopx is expressed in various hematopoietic cell subsets of C57Bl/6 mice (n = 3), including all three HSPC populations, but at very low levels in mesenchymal stem cells (MSCs). Expression is also abundant in mature leukocytes, particularly in T lymphocytes (CD3+) compared with B lymphocytes (B220+) or monocytes (CD11b+). Total RNA was isolated from BM and RT-PCR was carried out as described in the Experimental Procedures. Expression levels are shown relative to the expression of Hprt, as an endogenous control, in relative units (ru) with error bars representing SEM.

(C) As determined by relative GFP fluorescence intensity, Hopx expression was highest in the LSKCD150−CD48− population of HSPCs and especially abundant in CD3+ cells, consistent with the RT-PCR results. Fluorescence was quantitated by flow cytometry in different HSPC sub-populations and mature hematopoietic lineages in BM MNCs that were isolated from the femurs and tibias of 12-week-old genetically modified mice (n = 3) expressing GFP under the control of the endogenous Hopx promoter (Takeda et al., 2013). Data are expressed in a.u. Note the different x axes in the plots.