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Synthetic pre-miR-29b and scrambled negative control RNAs (pre-miR-control) were purchased from Ambion (Austin, TX, USA).
This same cell line was transfected with synthetic pre-miRNAs to stimulate overexpression of corresponding miR-200 members in vitro.
Synthetic pre-miR-10a, anti-miR-10a, pre-miR-control and anti-miR-control RNAs were purchased from Genepharma (Shanghai, China).
The introduction of synthetic pre-let-7 causes the cell cycle arrest in lung cancer cells [7].
To that end, we transfected lung fibroblasts HFL1 either with a synthetic pre-miR-155 or a "negative" pre-miRNA, used as a control.
Synthetic pre-miRNAs and nonspecific negative control (miRIDIAN microRNA Mimic Negative Control#1) were purchased from Dharmacon, Inc. (Lafayette, CO, USA).
To evaluate whether miR-155 can alter the expression of KGF, we cloned a fragment of 919 and 787 bp of the human and murine KGF 3'-UTR mRNA (NM_002009 and NM_008008) containing the two putative miRNA-binding sites into the psiCHEK™-2 vector (Fig. 3B) and transfected it into HEK 293 or NIH3T3 cells in the presence of either a human or murine synthetic pre-miR-155 analogue or a pre-miR-control.
Synthetic pre-miR-591 or pre-miR-106a (Ambion) of 20 60 nℳ was added to the above reactions.
For instance, we generated stable clones while previous studies performed transient transfections with synthetic pre-miR-143.
Synthetic pre-mir-16, anti-mir-16, and scrambled negative control RNAs were purchased from Ambion (Austin, TX, USA).
Synthetic pre-miR-141 and scrambled negative control RNA (pre-miR-control) were purchased from Ambion (Austin, TX, USA).
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