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Luciferase reporter assays using pGL2-Basic vector were performed in order to investigate the functional effect of mutations on DC-SIGNR promoter activity.
Transfections of myr-Akt1 (activated) or empty plasmid (vector) were performed with Lipofectamine 2000 (Invitrogen, 11668019) according to the protocol suggested by the manufacture.
The insertion of a complete attB adaptor and cloning into the pDONRTM201 vector were performed in accordance with the manufacturer's protocol to create an entry clone for use during the subsequent subcloning steps.
Control nucleofections using the empty vector were performed in all experiments.
Transfections with this plasmid and the empty vector were performed with Lipofectamin (Invitrogen GIBCO/Life Technologies, Carlsbad, CA, USA).
Expression trials using murine cDNA for USP18 cloned into pET15b or a pGEMEX vector were performed in E. coli Rosetta DE3).
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Large-scale production and purification of vectors were performed by Vector Gene Technology Company Ltd.
Next, a compensation of the feature vector was performed employing MMSE estimation of the log-spectrum vector.
For further optimization of cofactor regeneration, now a co-expression of CHMO and ADH in E. coli using a Duet™ vector was performed.
Lipofection of expression vector was performed as recommended in the supplier's protocol using FuGENE 6 (Roche, Basel, Switzerland).
Transmission electron microscopy examination of HEK293T cells transfected with MAVS or empty vector was performed at 48 hours post-transfection (Fig. 2D).
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