Ez magna g chromatin immunoprecipitation kit

Chromatin was immunoprecipitated according with the EZ-Magna G Chromatin Immunoprecipitation kit's (Millipore Darmstadt, GER) user guide. Occupancy was assessed by qRT-PCR for samples precipitated with a speci c antibody versus samples precipitated with the control immunoglobulin G (12-371, Millipore Darmstadt, GER) using the speci c primers indicated in the Additional le 3: Supplemental Table S2.
Human umbilical vein endothelial cell (HUVEC) tube formation assay HeLa cells with ectopic FOXM1A/B/C/D expression were cultured in complete medium for 12 h, and then serum-free medium for 24 h, after which point the supernatants were collected. Matrigel (BD Biosciences, Franklin Lakes, NJ) (50 μL) was pipetted into each well of a 96-well plate and polymerized for 30 min at 37℃ in incubator. HUVECs (4×10 4 cells) were suspended in 100 μL of the above supernatants and distributed into each well. After incubation for 5 h at 37℃, images were taken with a 100× and 400× phase contrast microscope. Each condition was assessed at least in triplicate in three independent experiments.