For siRNA‐KD experiments, H9c2 cells were seeded at 106 cells per 100 mm dish and transfected with 25 nM scrambled siRNA (Invitrogen AM4611), Rbfox2 siRNA (Invitrogen siRNA ID# s96620) or PTBP siRNAs (Qiagen cat# SI02649206 and SI04255146) using Lipofectamine RNAiMAX (Thermo Fisher Scientific). Cells were harvested 72 h post‐transfection for RNA or protein extraction. For rescue experiments, 3 × 106 H9c2 cells were transfected with eGFP (Sigma‐Aldrich), human GFP‐RBFOX2 (transcript variant 3) (Addgene, plasmid #63086) or empty vector (pcDNA 5) together with scrambled or Rbfox2 specific siRNAs using Neon Nucleofection System (Thermo Fisher Scientific) as described previously.38 (link) RNA was harvested 48 h post‐transfection.
Human gfp rbfox2 transcript variant 3
Manufactured by Addgene
Human GFP‐RBFOX2 (transcript variant 3) is a recombinant protein that contains the green fluorescent protein (GFP) fused to the RBFOX2 protein. RBFOX2 is a splicing factor that regulates alternative splicing. The fusion of GFP to RBFOX2 allows for the visualization and tracking of the RBFOX2 protein in cells.
Automatically generated - may contain errors
2 protocols using human gfp rbfox2 transcript variant 3
1
siRNA Knockdown and Rescue Experiments in H9c2 Cells
2
Rbfox2 and PTBP knockdown in H9c2 cells
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For siRNA-KD experiments, H9c2 cells were seeded at 10 6 cells per 100mm dish and transfected with 20nM scrambled siRNA (Invitrogen AM4611), Rbfox2 siRNA (Invitrogen siRNA ID# s96620) or PTBP siRNAs (Qiagen cat# SI02649206 and SI04255146) using Lipofectamine RNAiMAX (Thermofisher Scientific). Cells were harvested 72 hours posttransfection for RNA or protein extraction. For rescue experiments, 3X10 6 H9c2 cells were transfected with eGFP (Sigma-Aldrich), human GFP-RBFOX2 (transcript variant 3) (Addgene, plasmid #63086) or empty vector (pcDNA 5) together with scrambled or Rbfox2 specific siRNAs using Neon Nucleofection System (Thermofisher Scientific) as described previously (Verma et al. 2013) . RNA was harvested 48 hours post-transfection.
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