For siRNA-KD experiments, H9c2 cells were seeded at 106 cells per 100mm dish and transfected with scrambled siRNA (Thermofisher Scientific AM4611) or Rbfox2 siRNA (Thermofisher Scientific siRNA ID# s96620) at 20nM using Lipofectamine RNAiMAX (Thermofisher Scientific 13778150). Cells were harvested 72 hours post-transfection for RNA or protein extraction. For rescue experiments, 3X106 H9c2 cells were transfected with eGFP (Sigma-Aldrich), human GFP-RBFOX2 (Addgene, plasmid #63086), Flag-ANT1 (SinoBiological HG17969-NF), or empty vector (pcDNA 5) together with scrambled or Rbfox2 specific siRNAs by Neon Nucleofection System (Thermofisher Scientific) and harvested 48 hours post-transfection as we have described (Verma et al., 2013 (link)). pEGFP-RBFOX2 (transcript variant 3) was a gift from Nicolas Charlet-Berguerand (Addgene plasmid # 63086; http://addgene.org/63086 ; RRID:Addgene_63086).
Flag ant1
Manufactured by Sino Biological
Flag-ANT1 is a recombinant protein that consists of a protein of interest fused to an N-terminal FLAG tag. The FLAG tag is a widely used epitope tag that can be used for the detection and purification of the recombinant protein.
Automatically generated - may contain errors
2 protocols using flag ant1
1
siRNA-Mediated Rbfox2 Knockdown in H9c2 Cells
2
siRNA-Mediated Rbfox2 Knockdown in H9c2 Cells
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For siRNA-KD experiments, H9c2 cells were seeded at 106 cells per 100mm dish and transfected with scrambled siRNA (Thermofisher Scientific AM4611) or Rbfox2 siRNA (Thermofisher Scientific siRNA ID# s96620) at 20nM using Lipofectamine RNAiMAX (Thermofisher Scientific 13778150). Cells were harvested 72 hours post-transfection for RNA or protein extraction. For rescue experiments, 3X106 H9c2 cells were transfected with eGFP (Sigma-Aldrich), human GFP-RBFOX2 (Addgene, plasmid #63086), Flag-ANT1 (SinoBiological HG17969-NF), or empty vector (pcDNA 5) together with scrambled or Rbfox2 specific siRNAs by Neon Nucleofection System (Thermofisher Scientific) and harvested 48 hours post-transfection as we have described (Verma et al., 2013 (link)). pEGFP-RBFOX2 (transcript variant 3) was a gift from Nicolas Charlet-Berguerand (Addgene plasmid # 63086; http://addgene.org/63086 ; RRID:Addgene_63086).
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