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. 2014 Jan 9;124(2):656–669. doi: 10.1172/JCI70090

Figure 3. WT FHC and mutant FHC 222 similarly inhibit CXCR4 signaling in HOS cells.

Figure 3

(A) WT FHC, mutant FHC 222 FHC (Mut; non–iron-binding), or EV was transfected into HOS cells (left), and the specific inability of mutant FHC 222 to bind iron was confirmed by calcein assay (right), a measure of free intracellular iron levels. Calcein fluorescence was increased by treatment with DFO or by overexpressing WT FHC, but not mutant FHC 222. (B) Effects of these proteins on CXCL12 signaling were compared by treating cells with 20 nM CXCL12 for various times. WT FHC– and mutant FHC 222–expressing cells demonstrated similar reductions in CXCL12-induced Akt and CXCR4 activation (n = 4). *P < 0.05, #P < 0.01, §P < 0.001 versus empty vector. See complete unedited blots in the supplemental material.