Integrin engagement regulates monocyte differentiation through the forkhead transcription factor Foxp1
J. Clin. Invest. Can Shi, et al. 114:408 doi:10.1172/JCI21100 [
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Figure 6Effect of full-length
MFH/Foxp1 and amino- and carboxy-terminal deletion mutants on
c-fms promoter activity. (
A) Activity of the
c-fms promoter. To map the repression domain of
MFH/Foxp1, a series of NH
2- and COOH-terminal
MFH/Foxp1 deletion mutants were generated. Deletion mutants were designed on the basis of the predicted modular domain structure of
MFH/Foxp1 in order to establish the contribution of the glutamine-rich (Q-rich, amino acids 55–200), zinc-finger (Zn-finger, amino acids 308–331), and winged-helix/
forkhead DNA-binding (amino acids 465–536) regions to the repressor activity of
MFH/Foxp1. NIH 3T3 cells were transfected with 0.5 μg
c-fms reporter gene plasmid, 0.05 μg pCMV-β-gal, and expression plasmids (5.5 μg) for pcDNA3.1 (vector), full-length
MFH/Foxp1, or deletion mutants added to each well of a 12-well plate. Luciferase activities were determined, normalized on the basis of β-galactosidase activity, and plotted as percent promoter activity compared with that induced by treatment with vector alone. Triplicate determination of three to five independent experiments (mean ± SD) is shown. *
P < 0.01. (
B) Verification of
MFH/Foxp1 mutant expression. Western blot analysis of NIH 3T3 lysates verifies expression of
MFH/Foxp1 and NH
2- and COOH-terminal
MFH/Foxp1 deletion mutants: vector alone (lane 1), full-length
MFH/Foxp1 amino acids 1–677 (lane 2), 430–677 (lane 3), 224–677 (lane 4), 1–222 (lane 5), and 1–428 (lane 6).
