MCF-7/ADR cells were seeded in 96-well plates for 24 h until they reached 90–95% confluence at the time of transfection. The cells were co-transfected with the MDR1 promoter recombinant vector pGL3-basic-MDR1, and a control vector according to the manufacturer’s instructions (Promega Corporation, Madison, WI, USA). The cells were collected 48 h after transfection and analyzed using a Dual-Luciferase Reporter assay system (Promega Corporation).
Control vector
Manufactured by Promega
Sourced in United States
The Control vector is a laboratory tool used to provide a reference point for experimental analysis. It serves as a standard for comparison, allowing researchers to evaluate the performance and reliability of their experimental systems. The Control vector offers a consistent and well-characterized genetic sequence that can be used to validate experimental procedures and interpret results.
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Lab products found in correlation
5 protocols using control vector
1
MDR1 Promoter Activity Assay
2
Identification and Validation of miR-206 Binding in CCL2 3'UTR
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The 3′untranslated region (UTR) of CCL2 containing the putative binding site of miR-206 was identified by searching the online miRNA database (18 (link),19 (link)). The CCL2 3′UTR was amplified using PCR, and the PCR products were isolated using agarose gel (2%). The fragments were then purified and inserted into a pGL3-control vector (Promega Coproration, Madison, WI, USA). Mutagenesis was performed for the same site and inserted into the control vector (Promega Corporation) using PCR. The miR-206 primers were added into the reverse transcription system. The miRNA reverse transcription program is consistent with the common mRNA procedures (20 (link)).
3
Plasmid Transfection and siRNA Knockdown in CRC Cells
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pcDNA3.1‐PTEN plasmid or the control vector (Promega) were transfected into CRC cells using Lipofectamine 3000 (ThermoFisher Scientific) following instructions, respectively. For siRNA transfections, CRC Cells were transfected with 1.5 μg/ml of siRNA using Lipofectamine RNAiMAX (Invitrogen).
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pcDNA3.1‐PTEN plasmid or the control vector (Promega) were transfected into CRC cells using Lipofectamine 3000 (ThermoFisher Scientific) following instructions, respectively. For siRNA transfections, CRC Cells were transfected with 1.5 μg/ml of siRNA using Lipofectamine RNAiMAX (Invitrogen).
4
PTEN Regulation by miR-424 Interaction
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TargetScan data were utilized to predict the binding site between PTEN and miR-424. Subsequently, a luciferase reporter assay was performed. Wild-type and mutant reporters (PTEN-WT and PTEN-MUT), along with NC mimics and miR-424 mimics, were co-transfected into cells using Lipofectamine 2000. Dual-luciferase reporter assay was performed according to the instruction. PTEN-sh-mut (5'-CGCGGATCCAAATATTACATTA-AGGGTTAAGTGATTTGATTCAGTAATTGTGCTATTCCTA-CATGTGCTTTATAGATTAAAGAATTGATTTTTTGG-TACCCCG-3') were synthesized by Augct (Beijing, China). The PTEN 3'UTR fragments were inserted downstream of the control vector (Promega, USA) via Fse I and Xba I sites.
5
NF-κB Pathway Activation Assay
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HEK293T cells were transfected with NF-κB -luciferase reporter plasmid together with control vector (Promega, USA) as reference controls using Lipofectamine 3000 (Invitrogen, USA). 48 hours later, cells were subject to different treatments. The luciferase activity measurement was performed as described in Dual luciferase reporter assay kit (Promega, USA).
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