Sigenome mouse siesr1

In this series of experiments, we used siRNA to knockdown Esr1 in the presence and absence of estrogens to determine if Esr1 mediates estrogens’ effects on Vegfa. To do this, 3T3-L1 cells were transfected with siRNA targeting murine Esr1 (siGENOME Mouse siEsr1, GE Dharmacon, Lafayette, U.S.). As a control, cells were treated with an unrelated control siRNA (siGENOME non-targeting siRNA, GE Dharmacon), using Lipofectamine RNAi max transfection reagent (Thermo Fisher), according to the manufacturer’s instructions. 48h after siRNA transfection, cells were treated with estrogens and as indicated, and then lysed for RNA extraction.

In this series of experiments, we used siRNA to knockdown Esr-1 (ERα gene) in the presence and absence of estrogens to determine if ERα mediates estrogens’ effects on inflammation. To do this, 3T3-L1 cells were cultured in 12-well plates and transfected at day 10 of differentiation with siRNA targeting murine Esr-1 (siGENOME Mouse siEsr1, GE Dharmacon, Lafayette, USA, at the concentration of 100 nM). As a control, cells were treated with an unrelated control/scrambled sequence siRNA at the same concentration (siGENOME non-targeting siRNA, GE Dharmacon). Lipofectamine RNAi max was used as the transfection reagent (Thermo Fisher, at the concentration of 20 рM/μl), according to the manufacturer’s instructions. Seventy-two hours after siRNA transfection, cells were treated with estrogens and LPS as indicated, and then lysed for RNA extraction.