Cells were seeded in six-well plates and were incubated with antibiotic-free medium half an hour before transfection. si-FGG, si-STAT3, and non-targeting control siRNA (si-control) (Synbio Technologies, China) were transfected in Opti-MEM medium (Invitrogen) using X-tremeGENETM siRNA Transfection Reagent (Roche, Germany) for 6 h according to the manufacturer’s instructions. The transfection medium was then replace with normal medium and the cells were cultured for an additional 48 h for subsequent experiments.
X tremegenetm sirna transfection reagent
Manufactured by Roche
Sourced in Germany
The X-tremeGENE siRNA Transfection Reagent is a lipid-based reagent designed to facilitate the delivery of small interfering RNA (siRNA) into a variety of cell types for gene silencing applications. The reagent is formulated to promote efficient uptake and intracellular delivery of siRNA, enabling researchers to effectively study gene function.
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2 protocols using x tremegenetm sirna transfection reagent
1
siRNA Transfection in Cell Lines
2
Gene Knockdown by RNAi Oligos
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For knockdown of gene expression, sets of three synthesized duplex RNAi oligos targeting human TTC22, WTAP, YTHDF1, YTHDF2, YTHDF3 or YTHDC1 mRNA were purchased from Gene Pharma (Shanghai, China; Table S1). Two siRNAs for RPL4 were purchased from RuiBo Bio (stB0001897A for siRPL4#1 and stB0001897B for siRPL4#2; Guangzhou, China). The LV3(H1/GFP& Puro)-shWTAP and empty vector plasmids were also constructed by GenePharma (Shanghai, China). Lentivirus particles of pLV-hU6-shRPL4 and the negative control were from SyngenTech Co., Ltd (pHS-ASR-1219 and pHS-ASR-LW429, Beijing, China). Cells at 70-80% con uence were transfected with siRNAs using X-tremeGENETM siRNA Transfection Reagent or with plasmids using X-tremeGENE HP DNA Transfection Reagent (Roche, Mannheim, Germany) according to the manufacturer's instructions. Successful knockdown of the target gene was con rmed by Western blot and qRT-PCR analyses. Scrambled siRNA sequences were used as negative controls.
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