Computational protocol: Keap1-Independent Regulation of Nrf2 Activity by Protein Acetylation and a BET Bromodomain Protein

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Protocol publication

[…] dsRNAs (200–700 bp) were synthesized and purified following the protocol provided as described [] using ‘T7 RiboMAX Express RNAi System’ kit (Promega) and ‘RNeasy Kit’ (QIAGEN). Briefly, 1X106 S2 cells were bathed with 8μg dsRNAs and these cells were transfected with luciferase reporter plasmids using the calcium phosphate method 3 days after dsRNA treatment. Predesigned dsRNAs obtained through the E-RNAi webservice [] were used to knock down Fs(1)h, CncC, Keap1 whereas Fs(1)h-L was knocked down using the dsRNA described by Kockmann et al. []. dsRNA targeting the unique 3’ UTR region of Fs(1)h-S was designed by the ‘SnapDragon’ webservice. The sequences of primers used to generate amplicons for dsRNA synthesis are provided in Table B in . […]

Pipeline specifications

Software tools E-RNAi, SnapDragon
Application Non-coding RNA analysis
Organisms Drosophila melanogaster, Homo sapiens