Cells were interfered with p62-, LC3-36 (link), 37 (link) or ATG5-siRNA (Qiagen, Hilden, Germany) using the Oligofectamine method (Invitrogen, Carlsbad, CA, USA) to suppress the expression of endogenous genes. EGFP-siRNA38 (link) was used as a non-specific control. Cells were harvested 48 hours post-transfection and reduction of protein levels confirmed by Western blotting.
Oligofectamine method
Manufactured by Thermo Fisher Scientific
Sourced in Germany, United States
Oligofectamine is a transfection reagent used for the delivery of nucleic acids, such as plasmids or siRNA, into eukaryotic cells. It facilitates the efficient uptake of these molecules by the cells, enabling their use in various research applications.
Automatically generated - may contain errors
5 protocols using oligofectamine method
1
Endogenous Gene Silencing Techniques
2
Silencing VEGF-C and Akt for Cell Studies
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Small interfering RNA (siRNA) duplexes specific for VEGF-C or Akt were purchased from Santa Cruz Biotechnology (Santa Cruz, Calif.); control siRNAs with sequences that do not target any gene product were obtained from Invitrogen (Carlsbad, Calif.). Cells were transfected with 25 nM VEGF-C siRNA, Akt siRNA, or control siRNA in serum-free Opti-MEM using the oligofectamine method (Invitrogen) for 1 h at 37 °C. After changing the culture medium, cells were cultured for 24 h at 37 °C prior to further experiments.
3
Knockdown of Galectin-3 in Cell Lines
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Small interfering RNA (siRNA) duplexes were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). The targeted siRNAs for human galectin-3, mouse galectin-3, and control siRNA were sc-155994, sc-35443 and sc-37007, respectively. The THP-1 and Raw 264.7 cells were transfected with siRNA at a concentration of 25 nM in serum-free Opti-MEM using the Oligofectamine method (Invitrogen, Carlsbad, CA, USA).
4
Suppressing Endogenous Genes via RNAi
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Cells were interfered with FBXW7α-, CDH1-, ATM- or ATR-siRNA [37 (link), 61 (link), 62 (link)] using the Oligofectamine method (Invitrogen) to suppress the expression of endogenous genes. EGFP-siRNA [61 (link)] was used as a non-specific control. Cells were harvested 48h post-transfection and reduction of protein levels confirmed by Western blotting.
Half-life experiments were performed by interfering cells with FBXW7α-siRNA or EGFP-siRNA and, 48h later, adding cycloheximide to the medium. Cells were harvested at the indicated times.
Half-life experiments were performed by interfering cells with FBXW7α-siRNA or EGFP-siRNA and, 48h later, adding cycloheximide to the medium. Cells were harvested at the indicated times.
5
Endogenous Gene Silencing Techniques
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Cells were interfered with βTrCP1/2-, GSK3β- or PKR-siRNA [37 (link)-39 (link)] using the Oligofectamine method (Invitrogen) to suppress the expression of endogenous genes. EGFP-siRNA [38 (link)] was used as a non-specific control. Cells were harvested 48h post-transfection and reduction of protein levels confirmed by Western blotting.
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