miR-130a down-regulates HOXA5 expression. (A) Mitogens down-regulate HOXA5 expression in HUVECs. HUVECs were transfected with full-length HOXA5 (pcDNA3.1-HOXA5), incubated overnight in either 0.1% FBS or 2.0% FBS, and then subjected to immunofluorescence with anti-Flag antibody. The photomicrograph was taken at 400× magnification; fluorophores and confocal microscope are described in “Immunofluorescence.” As with GAX and consistent with previously reported work,18 serum resulted in the down-regulation of HOXA5 expression.(B) miR-130a down-regulates HOXA5 expression. HUVECs were cotransfected with either pcDNA3.1 or pcDNA3.1-miR-130a plus pcDNA3.1-HOXA5, incubated overnight, and then harvested for Western blot with anti-Flag antibody. miR-130a strongly down-regulates HOXA5 expression. (C) Quantification. Western blots were subjected to densitometry and HOXA5 band densities normalized to those of the corresponding α-tubulin bands. Error bars represnt SEM.
