RNA pull‐down assay was conducted according to the previous reports.
30 (link) Briefly, the LINC01128 overexpression vector was designed by Sangon Biotech to serve as a template to amplify the LINC01128‐corresponding antisense and sense strands using the T7 promoter‐containing primers. Biotinylated LINC01128 was synthesised by the TranscriptAid T7 High Yield Transcription Kit (Thermo Fisher Scientific), which was subsequently incubated with streptavidin‐coated magnetic beads. The RNA pull‐down assay was conducted using the Magnetic RNA‐Protein Pull‐Down Kit (Pierce 20164, Thermo Fisher Scientific). Subsequently, the precipitated proteins were employed for silver staining, western blotting and mass spectrometry. The RNA–protein interaction probabilities generated byhttp://pridb.gdcb.iastate.edu/RPISeq/about.php (RNA–Protein Interaction Prediction, RPISeq) ranged from 0 to 1. Reaction probabilities were represented as values of random forest (RF) and support vector machine (SVM), and only those values where both RF >.5 and SVM >.5 were identified as ‘positive’, displaying that the possible interaction of protein with RNA.
30 (link) Briefly, the LINC01128 overexpression vector was designed by Sangon Biotech to serve as a template to amplify the LINC01128‐corresponding antisense and sense strands using the T7 promoter‐containing primers. Biotinylated LINC01128 was synthesised by the TranscriptAid T7 High Yield Transcription Kit (Thermo Fisher Scientific), which was subsequently incubated with streptavidin‐coated magnetic beads. The RNA pull‐down assay was conducted using the Magnetic RNA‐Protein Pull‐Down Kit (Pierce 20164, Thermo Fisher Scientific). Subsequently, the precipitated proteins were employed for silver staining, western blotting and mass spectrometry. The RNA–protein interaction probabilities generated by
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