miR-150 Targets Cxcr4 to Regulate MNC Mobilization incubating the complex at room temperature, the DNA-Endo- Fectine complex was added in the dish which 293Ta cells were plated in DMEM with 10% FBS and incubated in 5% CO2 at 37uC overnight. The culture medium was replaced with fresh DMEM with 5% FBS and 1/500 volume of the TiterBoost reagent to the culture medium and continued to be incubated. The virus pseudovirus-containing culture medium was collected 48 hr post transfection and concentrated after filtration. For the transduction of BM-MNCs with lentivirus, 106106 of MNCs was plated and 20 ml of virus suspension was added. The cells were placed for 2 hr at 4uC and then transferred to the plate in a 5% CO2 at 37uC for 48 h. Figure 1. AMI increases the numbers of BM-MNCs and CXCR4 positive MNCs. (A) MNCs were isolated at 1, 3 and 5 days after left anterior descending artery ligation (LAD) and analyzed for molecular characterization. (B) The number of PB-MNCs decreased at one day after LAD ligation and increased gradually up to 5 days (n$10 in each groups). (C) The percentage of CXCR4 positive cells in MNCs increased in both PB and BM after LAD ligation as compared to control. (n = 8 in control group and n = 6 in AMI group). (D) Densitometric analysis of CXCR4 positive MNCs counted after LAD ligation. doi:10.1371/journal.pone.0023114.g001 PLoS ONE | www.plosone.org 3 October 2011 | Volume 6 | Issue 10 | e23114