Protocols using G-CSF to mobilize stem cells and echocardiographic assessment of cardiac function in mice. (A) Animals were divided into four groups for 4 weeks of iron intra- peritoneally injection (10 mg/25 gm body weight of mouse per day for 5 days/week) or dextran injection as shown in the protocols. G-CSF (100 μg/kg/day subcutaneous injection) or saline was given for 5 days in the second week as shown. I+G; iron plus G-CSF treatment. (B) Different dosages of G-CSF were given to mice with blood c-kit and CD45 examined by flow cytometry analysis. (C) Representative echocardiograms of mitral-valve-flows Doppler mapping (E and A waves) in each experimental group at end of the second and fourth week, respectively. Decreased E: A wave ratio showing diastolic dysfunction in the I+G group. E wave and A wave, indicating LV early-filling wave and filling from atrial contraction, respectively. (D) Representative 2D echocardiogram of long axis view revealed intra-cardiac mass (arrow) in the apex region of the left ventricle in the I+G group at 4th week exam.