Potentials of GM-IMs to differentiate to M1 or M2 macrophages.

<div><p>GM-IMs or PMφ were stimulated with 20 ng/mL IFNγ, 20 ng/mL IL-4 and 20 ng/mL IL-13 or ong µg/mL LPS for 24 hr.</p> <p>(A) Phase contrast photographs of GM-IMs after stimulation. Scale Bar: 100 µm.</p> <p>(B) After stimulation by IFNγ for 24hr, expression of M1 marker genes was analyzed by RT-PCR.</p> <p>(C) After stimulation by LPS, RNA was extracted and expression of M1 marker genes was analyzed by RT-PCR.</p> <p>(D) After stimulation by IL-4 and IL-13, expression of M2 marker genes was analyzed by RT-PCR.</p> <p>(E) GM-IMs or BMMφ (1 x 10⁶) were cultured in six-well plastic plates with RPMI-1640 containing 10% FBS and 3% GM-CSF-CM. They were stimulated with one µg/mL LPS for indicated times. Supernatants were taken and cytokine expression of IL-6, IL-1β or TNFα was analyzed by ELISA. Data shown are the mean ratios ± SE of three separate experiments.</p> <p>(F) The dorsal skin of two-month old female C57BL/6 mice (n=4) was punctured through two layers of skin with a sterile disposable three mm biopsy punch (Kai Industries). GM-IMs (3 x 10⁴) were directly injected into each wound on the right side. PBS was dropped on the other side as a control. The changes in the percentages of wound areas at each time point were compared to the initial wound area. Data shown are the mean ratios ± SE of three independent experiments. P value: * < 0.05, **<0.01, ***<0.001, GM-IMs injected area versus PBS injected area.</p> <p>Data are representative of three independent experiments with similar results (A-D).</p>