The miRNA mimic, miRNA inhibitor, STAT3 siRNA, and scrambled siRNA were synthesized by RiBoBio (China). The oligonucleotide sequences were listed in Table S1 (Additional file 1 : Table S1). The miRNA overexpression vector pCMV-MIR519A (MI0003182) and the empty vector control were obtained from OriGene (Rockville, MD, USA). The miR-519a sponge and empty vector control were purchased from GeneChem (China). Transfections were performed using Lipofectamine 2000 reagent (cat. no. 11668-019; Invitrogen, Carlsbad, CA, USA) according to the manufacturer’s instructions.
Empty vector control
Manufactured by Genechem
Sourced in China
The empty vector control is a plasmid that does not contain any specific gene of interest. It is commonly used as a negative control in experiments involving gene expression or gene manipulation.
Automatically generated - may contain errors
Lab products found in correlation
Stat3 sirna, by RiboBio (1 mentions)
Mirna inhibitor, by RiboBio (1 mentions)
Mirna mimic, by RiboBio (1 mentions)
Empty vector control, by OriGene (1 mentions)
Scrambled sirna, by RiboBio (1 mentions)
Lipofectamine 3000, by Thermo Fisher Scientific (1 mentions)
Nc mimics inhibitor, by RiboBio (1 mentions)
Lipofectamine 2000, by Genechem (1 mentions)
4 protocols using empty vector control
1
Transfection of miRNA and siRNA
2
Silencing SNHG22 and Upregulating SUDS3 in Breast Cancer
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The specific shRNAs against SNHG22 and corresponding negative control shRNAs, as well as the pcDNA3.1 vector containing SUDS3 and the empty vector (control), were all available from Genechem (Shanghai, China). Besides, the miR-324-3p mimics/inhibitor and NC mimics/inhibitor were synthesized by Ribobio (Guangzhou, China). MDA-MB-231 and MDA-MB-468 cells prepared in 24-well plates were subjected to transfection with appropriate plasmids above in the presence of Lipofectamine 3000 (Invitrogen). After 48 h of transfections, cells were collected for further use in following experiments.
3
Plasmid Transfection Enhances Anoikis
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For plasmid transfection, Bel-7402 cells were cultured on 6-well plate to 90–95% confluence, and 4.0 μg recombinant human PTK2 eukaryotic expression plasmid or control empty vector (Genechem, Shanghai, China) were introduced into the cells by Lipofectamine™ 2000 according to the manufacturer’s recommendations. After 24 h of transfection, cells were subjected to suspension-culture, YGJDSJ (200 μg/ml) treatment for 24 h, western blot and anoikis assay.
4
Lentiviral Knockdown and Overexpression of PER2
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For PER2 knockdown, the lentivirus expression vectors PER2-sh1, PER2-sh2 and control (empty vector) were purchased from GeneChem (Shanghai, China). For PER2 overexpression (PER2-OE), the lentivirus expression vectors pCDNA3.1-PER2-3×FLAG and the control vector were purchased from Miaolingbio (Wuhan, China). Control vectors or PER2 knockdown/overexpression vectors, pMD2G and psPAX2 vectors were co-transfected into 293 T cells with PEI according to the manufacturer’s protocol. After 48 h, the supernatant in 293 T cell dishes was collected for lentivirus. Human DPSCs were transfected with the corresponding lentivirus particles and renamed based on the lentiviral vectors used, and the final efficiency of knockdown or overexpression of PER2 was evaluated by qRT-PCR and Western blot analysis.
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