MirScript miRNA 142-3p mimic and inhibitor were purchased from Qiagen. AllStars negative mimic (Qiagen) was used as control. Gene specific and control siRNA for PKCα knockdown were purchased from Sigma-Aldrich. Transient transfections were performed using Lipofectamine 2000 (Life Technologies) according to manufacturer’s instructions. Cells were transfected with mimics or inhibitors at a final concentration of 100 nM. Red siGLO oligonucleotides (Thermo Fisher Scientific) were used to determine transfection efficiency (S2 Table). Viability was assessed by flow cytometry (S3 Table).
Allstars negative mimic
Manufactured by Qiagen
AllStars negative mimics are a set of synthetic RNA molecules designed to serve as negative controls in RNAi experiments. They are engineered to not target any known gene sequences, allowing researchers to evaluate the specificity and effectiveness of their RNAi reagents.
Automatically generated - may contain errors
Lab products found in correlation
Lipofectamine 2000, by Thermo Fisher Scientific (4 mentions)
Red siglo oligos, by Thermo Fisher Scientific (3 mentions)
Celltiter 96 aqueous cell proliferation assay kit, by Promega (2 mentions)
Miscript mirna mimics and inhibitors, by Qiagen (2 mentions)
Lipofectamine 2000 reagent, by Thermo Fisher Scientific (1 mentions)
5 protocols using allstars negative mimic
1
Transfection of miRNA mimic and inhibitor
2
Transfection of miRNA Mimics and siRNAs
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miRNA mimics and siRNAs were purchased from Shanghai GenePharma. AllStars negative mimic (Qiagen) or Silencer (Life Technologies) were used as negative controls. Cells were transfected with siRNA or micrRNA mimic at a final concentration of 100 pM using Lipofectamine 2000 reagent (Invitrogen) according to the manufacturer's instructions. Sequences for siRNAs and miRNA mimics are presented in Supplementary Table 7 .
3
miRNA Mimic Transfection in Differentiated mD-Mφ
4
miRNA Mimic Transfection in Differentiated mD-Mφ
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MiScript miRNA mimics and inhibitors were purchased from Qiagen. AllStars negative mimics (Qiagen) were used as controls. Transient transfections were performed using Lipofectamine 2000 (Life Technologies) according to manufacturer’s instructions. Day 7 differentiated mD-Mφ were transfected at a final concentration of 50 nM. Red siGLO oligos (Thermo Fisher Scientific, Waltham, MA, USA) were used to confirm successful transfection, with transfection efficiency being greater than 90% as previously reported [9 (link)]. Cell viability was assessed using the CellTiter 96 AQueous Cell Proliferation Assay Kit (Promega, Madison, WI, USA). Viability of all cell culture samples was analyzed 2 hours prior to harvesting of cells/supernatant for subsequent experimental analysis.
5
MiRNA Mimics and Inhibitor Transfection
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