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Duplexed RNA oligonucleotides for rat EpoR and GATA1 (siGENOME SMARTpool TM) were synthesized by Thermo Fisher Scientific Inc. H19-7 cells were transfected with 40 pmol of GATA1-specific siRNAs (siGATA1), 40 pmol of EpoR-specific siRNAs (siEpoR) or scrambled siRNA (Sc) using transfection reagents (TransIT-TKO, Mirus) according to the manufacturer’s protocols. After transfection, the cells were then incubated for 48 hours prior to their use in further investigations. The efficiency of the EpoR and GATA1 protein silencing was evaluated by immunoblotting to optimize the experimental conditions (siRNA dose and time after transfection) following the previous studies (Rujitanaroj et al. 2013). |
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Protocol tips |
Duplexed RNA oligonucleotides for rat EpoR and GATA1 (siGENOME SMARTpool TM) were synthesized by Thermo Fisher Scientific Inc. H19-7 cells were transfected with 40 pmol of GATA1-specific siRNAs (siGATA1), 40 pmol of EpoR-specific siRNAs (siEpoR) or scrambled siRNA (Sc) using transfection reagents (TransIT-TKO, Mirus) according to the manufacturer’s protocols. After transfection, the cells were then incubated for 48 hours prior to their use in further investigations. The efficiency of the EpoR and GATA1 protein silencing was evaluated by immunoblotting to optimize the experimental conditions (siRNA dose and time after transfection) following the previous studies (Rujitanaroj et al. 2013). |
Publication protocol
Duplexed RNA oligonucleotides for rat EpoR and GATA1 (siGENOME SMARTpool TM) were synthesized by Thermo Fisher Scientific Inc. H19-7 cells were transfected with 40 pmol of GATA1-specific siRNAs (siGATA1), 40 pmol of EpoR-specific siRNAs (siEpoR) or scrambled siRNA (Sc) using transfection reagents (TransIT-TKO, Mirus) according to the manufacturer’s protocols. After transfection, the cells were then incubated for 48 hours prior to their use in further investigations. The efficiency of the EpoR and GATA1 protein silencing was evaluated by immunoblotting to optimize the experimental conditions (siRNA dose and time after transfection) following the previous studies (Rujitanaroj et al. 2013).
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