Macrophage colony stimulation factor m csf

17β-estradiol, Cat # E2257; progesterone, Cat # P7556; mifepristone, Cat# M8046; and tamoxifen, Cat # T5648 were purchased from Sigma Chemical Company, St. Louis, MO, USA. Macrophage colony stimulation factor (M-CSF), Cat # PHC2044 was purchased from Thermo Fisher Scientific, Waltham, MA, USA.

17β-estradiol, Cat # E2257 and progesterone, Cat # P7556 were purchased from Sigma Chemical Company, St. Louis, MO, USA. Macrophage colony stimulation factor (M-CSF), Cat # PHC2044 was purchased from Thermo Fisher Scientific, Waltham, MA, USA. SLPI, Cat # 1274-PI-100 and SERPIN C1, Cat # 1267-PI-010 were purchased from R&D Systems, Minneapolis, MN, USA.

Murine VSMCs were isolated from the aorta by enzyme isolation as previously described (27 (link)). Cultures were trypsinized, followed by the addition of CD31-conjugated Dynabeads (Pharmingen) to the cell suspension and magnetic separation of endothelial cells (ECs) from VSMCs. Selectivity of recovered VSMC populations was highly specific based on detection of α-smooth muscle actin protein by FACS analysis in the VSMC cultures (Supplementary Figure 1). VSMCs at passage 2 to 10 were used for experiments. Murine VSMCs were grown in Dulbecco's modified Eagle's medium (Thermo Fisher Scientific Inc., Waltham, MA, USA) supplemented with 10% fetal bovine serum (FBS; Thermo Fisher Scientific), penicillin (100 U/ml; Invitrogen Life Technologies, Carlsbad, CA), and streptomycin sulfate (Invitrogen Life Technologies). Mouse bone marrow cells were prepared from the femur and tibia of 6-week old female mice and cultured with 30 ng/ml macrophage colony stimulation factor (M-CSF) (Peprotech, Rocky Hill, NJ, USA) for 3 days to prepare bone marrow-derived macrophages (BMMs) as previously described (28 (link)). BMMs were grown in Minimal Essential Media (Thermo Fisher Scientific) supplemented with 10% FBS and 30 ng/ml M-CSF.