Figure 3.

miR‐574‐5p regulated apoptosis and mitogen‐activated protein kinase pathways via MAP3K9 in goat mammary epithelial cells (GMECs). (a) The apoptosis of GMECs transfected with miR‐574‐5p mimics, inhibitors, si‐MAP3K9 or pcDNA3.1‐MAP3K9 expression vectors or cotransfected MAP3K9 with miR‐574‐5p for 24 hr. The histogram showed the apoptotic cell percentage detected by flow cytometry method, and error bars denote mean ± SD. After 48 hr transfection, (b) western blot analysis was used to detect the expression of Bcl‐2 and Bax in GMECs. (c, d) Cytosolic proteins were analyzed by western blot for extracellular‐signal‐regulated kinase, p38, c‐Jun N‐terminal kinase, MAP2K4, MAP2K7, c‐Jun, RSK1, Bad, and cyclin D1. (e) GMECs were treated with 5 μM anisomycin (AN) and honokiol (HO) in 24 and 48 hr. The expression of Bcl‐2 and Bax was analyzed by western blot. β‐Actin was detected as a loading control. *p < .05; **p < .01