Silencer select_FGG siRNA

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	A human hepatocyte cell line, HepG2 cells, was cultured in Dulbecco's modified Eagle's medium (DMEM; GIBCO, Grand Island, NY, USA) containing 10% fetal bovine serum (FBS; HyClone Laboratories, Inc Logan, UT, USA), 50 U/ml penicillin, and 50 ?g/ml streptomycin (GIBCO) in a 5% CO2 incubator at 37 °C. Silencer Select Pre-designed FGG siRNA (catalog no. 4392420, ID s5179)and Negative Control #1 siRNA (catalog no. 4390843) were obtained from Ambion (Austin, TX, USA). Each siRNA was reversely transfected into HepG2 cells in technical triplicates in 3 independent experiments. Briefly, each siRNA was mixed with 100 ?l OPTI-MEM I Reduced Serum Medium (GIBCO) in a 24-well tissue culture plate, and 1 ?l Lipofectamine RNAiMAX (Invitrogen, Carlsbad, CA, USA) was added to each well, mixed gently, and incubated for 20 minutes at room temperature according to the manufacturer's protocol. Then, 500 ?l HepG2 cells suspension containing 5×104 cells was added to the mixture of siRNA reagent. The final concentrations of FGA, FGB, and FGG siRNA varied between 1.0 and 30 nM, negative control siRNA was used at 10 nM, and the absence of siRNA was used as a non-transfected control. To obtain a nearly 50% decrease of FGA, FGB, or FGG mRNA expression in each experiment, siRNA concentrations varied from 1.0 to 2.0 nM.

Protocol tips

A human hepatocyte cell line, HepG2 cells, was cultured in Dulbecco's modified Eagle's medium (DMEM; GIBCO, Grand Island, NY, USA) containing 10% fetal bovine serum (FBS; HyClone Laboratories, Inc Logan, UT, USA), 50 U/ml penicillin, and 50 ?g/ml streptomycin (GIBCO) in a 5% CO2 incubator at 37 °C. Silencer Select Pre-designed FGG siRNA (catalog no. 4392420, ID s5179)and Negative Control #1 siRNA (catalog no. 4390843) were obtained from Ambion (Austin, TX, USA). Each siRNA was reversely transfected into HepG2 cells in technical triplicates in 3 independent experiments. Briefly, each siRNA was mixed with 100 ?l OPTI-MEM I Reduced Serum Medium (GIBCO) in a 24-well tissue culture plate, and 1 ?l Lipofectamine RNAiMAX (Invitrogen, Carlsbad, CA, USA) was added to each well, mixed gently, and incubated for 20 minutes at room temperature according to the manufacturer's protocol. Then, 500 ?l HepG2 cells suspension containing 5×104 cells was added to the mixture of siRNA reagent. The final concentrations of FGA, FGB, and FGG siRNA varied between 1.0 and 30 nM, negative control siRNA was used at 10 nM, and the absence of siRNA was used as a non-transfected control. To obtain a nearly 50% decrease of FGA, FGB, or FGG mRNA expression in each experiment, siRNA concentrations varied from 1.0 to 2.0 nM.

Publication protocol

A human hepatocyte cell line, HepG2 cells, was cultured in Dulbecco's modified Eagle's medium (DMEM; GIBCO, Grand Island, NY, USA) containing 10% fetal bovine serum (FBS; HyClone Laboratories, Inc Logan, UT, USA), 50 U/ml penicillin, and 50 ?g/ml streptomycin (GIBCO) in a 5% CO2 incubator at 37 °C. Silencer Select Pre-designed FGG siRNA (catalog no. 4392420, ID s5179)and Negative Control #1 siRNA (catalog no. 4390843) were obtained from Ambion (Austin, TX, USA). Each siRNA was reversely transfected into HepG2 cells in technical triplicates in 3 independent experiments. Briefly, each siRNA was mixed with 100 ?l OPTI-MEM I Reduced Serum Medium (GIBCO) in a 24-well tissue culture plate, and 1 ?l Lipofectamine RNAiMAX (Invitrogen, Carlsbad, CA, USA) was added to each well, mixed gently, and incubated for 20 minutes at room temperature according to the manufacturer's protocol. Then, 500 ?l HepG2 cells suspension containing 5×104 cells was added to the mixture of siRNA reagent. The final concentrations of FGA, FGB, and FGG siRNA varied between 1.0 and 30 nM, negative control siRNA was used at 10 nM, and the absence of siRNA was used as a non-transfected control. To obtain a nearly 50% decrease of FGA, FGB, or FGG mRNA expression in each experiment, siRNA concentrations varied from 1.0 to 2.0 nM.

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