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).
Human gfp rbfox2
Manufactured by Addgene
The Human GFP-RBFOX2 is a fluorescent protein-based construct designed to visualize the localization and expression of the RBFOX2 protein in human cells. RBFOX2 is an important RNA-binding protein involved in regulating alternative splicing. The GFP (Green Fluorescent Protein) tag allows for the direct observation of the RBFOX2 protein within the cellular environment using fluorescence microscopy techniques.
Automatically generated - may contain errors
2 protocols using human gfp rbfox2
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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