6 × 104 MDA-MB-231 cells were seeded in each well of a 24-well plate. The following day, cells were transfected with 200 ng of pCMV6-AC-GFP empty vector control (Origene) or CORO1C ORF construct (pCMV6-AC-CORO1C-GFP) (Origene) with 1μl of Lipofectamine 3000 and 0.8μl of P3000 (Invitrogen), according to the manufacturer’s protocol and incubated overnight. Subsequently, the cells were transfected with siNeg or siYB-1 siRNA to a final concentration of 20 nM per well using DharmaFECT (GE Dharmacon) as described above. The cells were incubated for an additional of 48 h before seeding for migration or invasion assay.
Pcmv6 ac gfp empty vector control
Manufactured by OriGene
Sourced in United States
The PCMV6-AC-GFP empty vector control is a plasmid designed for use as a control in gene expression studies. It contains the cytomegalovirus (CMV) promoter and a green fluorescent protein (GFP) marker, but no insert gene. This vector can be used to assess background expression levels and transfection efficiency in cell-based experiments.
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2 protocols using pcmv6 ac gfp empty vector control
1
Overexpression and Knockdown of CORO1C in MDA-MB-231 Cells
2
Overexpression of YBX1 in MDA-MB-231 Cells
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A cell density of 2.5 × 105 MDA-MB-231 cells was plated in each well of a 6-well plate, a day prior to plasmid transfection. The following day, the cells were transfected with 2 μg of pCMV6-AC-YBX1-GFP overexpression plasmid which encodes the ORF of YBX1 gene (Origene, Rockville, MD, USA) or the pCMV6-AC-GFP empty vector control (Origene) using 6 μl of TurboFectin 8.0 (Origene) per well. The transfected cells were then incubated for 72 h for protein isolation. Experiments were performed in triplicate wells for each plasmid.
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