Following the manufacturer's instructions, transfection was performed by using Lipofectamine 2000 (Invitrogen) with EphA2-specific siRNA (5′-AATGACATGCCGATCTACATG-3′) and the scramble sequence siRNA (GeneChem, Shanghai, China).
The sequences for hsa-miR-520d-5p hsa-miR-520d-5p hsa-miR-520d-5p hsa-miR-520d-5pandCCUUUC-3′ and 5′-GAGUCCGCCtheATAGACAA-3′, rescrambledy.
The scrambled sequence is shR-Scr, GCGATGCTCTAAGGTTCTATCAA.
Plakoglobin and γ-catenin expression were reduced by more than 80% in all knockdown cell lines used but were unaltered after transfection with the scrambled sequence.
As hypothesized, subjects' perception was biased by predictable sequences compared with the scrambled sequences.
To produce T47D and MCF7 cells with reduced levels of plakoglobin, pre-miRNA expression cassettes were designed to target the plakoglobin gene, and control cells were produced by using scramble sequence.
Eca109 cells were maintained in DMEM (Gibco) supplemented with 10% FBS (Gibco) in a 5% CO2 humidified incubator at 37°C and transfected with miR-21 mimics, miR-21 inhibitor, and scramble sequence by Lipofectamine 2000 in Eca109 cells as described previously [ 16].
This protein is the same length as K1009X but has lost the last few amino acid residues of the PEHE motif followed by scrambled sequence, whereas K1009X carries a wild-type PEHE domain.
The RNAi sequence for HPV16E7 (GCT TCG GTT GTG CGT ACA A) was identified by using the manufacturer's RNAi Designer programme, and the negative control having no homology with human genome was created by a scrambled sequence (TTC TCC GAA CGT GTC ACG T).
Non-targeting siRNA (NT-siRNA) was synthesized by a scrambled sequence of nucleotides as a control siRNA.
