siRNA transfection

HY Hironobu Yamashita
YK Yuka I. Kawasawa
LS Lauren Shuman
ZZ Zongyu Zheng
TT Truc Tran
VW Vonn Walter
JW Joshua I. Warrick
GC Guoli Chen
HA Hikmat Al-Ahmadie
MK Matthew Kaag
PW Pak Kin Wong
JR Jay D. Raman
DD David J. DeGraff
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siRNA transfection was performed using a reverse transfection method with lipofectamine 3000 (Thermo Fisher, Waltham, MA). Non-targeting (Scrambled; D-001810-01-05), TFAP2A (L-006348-02-0005), TFAP2C (L-005238-00-0005), and PPARG (L-003436-00) siRNAs were obtained from Dharmacon (Lafayette, CO). siRNA and lipofectamine 3000 were added to separate aliquots of OPTi-MEM medium, respectively, and incubated for 5 min. Subsequently, siRNA and lipofectamine 3000 in OPTi-MEM were mixed and incubated for 20 min. After incubation, siRNA-lipofectamine 3000 complex was added to 2 ml cell suspension containing 4 × 105 target cells per well in a 6-well plate.

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