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GLUT1 knockdown

QW Qian Wang
LN Lulingxiao Nie
PZ Pengfei Zhao
XZ Xinyi Zhou
YD Yi Ding
QC Qianming Chen
QW Qi Wang
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Small interfering RNA (siRNA) targeting mouse GLUT1 designed according to reference20 (5′-GGAATTCAATGCTGATGATGA-3′, 5′-TCATCATCAGCATTGAATTCC-3′, GenePharma Co.) was used to block GLUT1 in BMDM. Non-targeted siRNA as a negative control (5′-TTCTCCGAACGTGTCACGT-3′, 5′-ACGTGACACGTTCGGAGAA-3′) was used in wells not transfected with GLUT1 siRNA. Normal saline-treated with diethyl pyrocarbonate (Sigma-Aldrich Corp. Ltd., MO, USA.) was used to dissolve siRNA to reach a 20 μmol·L−1 concentration. The transfection mixture per well was prepared using 5 μL Endofectin (GeneCopoeia Inc., USA) and 5 μL GLUT1 siRNA or noncoding siRNA. After transfection for 48 h, the GLUT1−/− BMDM was cultured as described above.

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