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Dicer1

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DICER1 is a key enzyme involved in the processing of microRNA (miRNA) precursors. It functions to cleave double-stranded RNA into short, double-stranded fragments, which are then further processed into mature miRNAs that regulate gene expression.

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Lab products found in correlation

β actin, by Merck Group (3 mentions) Cyclin d1, by Cell Signaling Technology (1 mentions) Anti cd44 antibody, by Cell Signaling Technology (1 mentions) Gadph, by Cell Signaling Technology (1 mentions) Vinculin, by Cell Signaling Technology (1 mentions) Ab108398, by Abcam (1 mentions) Ab14140, by Abcam (1 mentions) Un scan it graph digitizer software 7, by Silk Scientific (1 mentions) Pvdf membrane, by Bio-Rad (1 mentions) Ripa buffer, by Merck Group (1 mentions) Image station 4000r pro, by Carestream (1 mentions) Dc protein assay, by Bio-Rad (1 mentions) Premade protease and phosphatase cocktail, by Thermo Fisher Scientific (1 mentions) Ecl system, by Cytiva (1 mentions) Drosha, by Cell Signaling Technology (1 mentions) Polyvinylidene fluoride membrane, by Merck Group (1 mentions) Cdp star reagent, by Roche (1 mentions) Quantity one densitometric software, by Bio-Rad (1 mentions)

4 protocols using dicer1

1

Comprehensive Molecular Profiling of Cells

Cited 1 time
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Western blotting and Immunohistochemically staining were performed as previously described [35 (link)]. The primary antibodies used in WB concludes of ESR1 (ab108398, 1:2000, Abcam), cyclin d1(#2978, 1:2000, Cell Signaling Technology), DICER1 (#3363, 1:2000, Cell Signaling Technology), NICD (#2421, 1:1000, Cell Signaling Technology), Vinculin (#13901, 1:5000, Cell Signaling Technology), NUMB (ab14140, 1:2000, Abcam), GADPH (#5174, 1:5000, Cell Signaling Technology). For immunobiological assay, the slides were incubated with anti-CD44 antibody (#3570, 1:500, Cell Signaling Technology), ALDH1 antibody (#54135, 1:500, Cell Signaling Technology) and NUMB antibody (ab14140, 1:1000, Abcam).
Xiao G., Li X., Li G., Zhang B., Xu C., Qin S., Du N., Wang J., Tang S.C., Zhang J., Ren H., Chen K, & Sun X. (2017). MiR-129 blocks estrogen induction of NOTCH signaling activity in breast cancer stem-like cells. Oncotarget, 8(61), 103261-103273.
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2

Western Blot Analysis of Protein Expression

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Whole cell extracts (WCE) were prepared in RIPA buffer (Sigma) with added phosphatase and complete protease inhibitors (Roche). Protein concentration was determined using the Bio-Rad DC protein assay (Bio-Rad). Where indicated, 10 or 15 μg of protein was boiled for 5 min, electrophoresed on 10% SDS-PAGE gels and electroblotted on PVDF membranes (Bio-Rad) for western blotting with the following antibodies: ERα (G-20, Santa Cruz Biotechnology, Santa Cruz, CA, USA), DICER1, PTEN (Cell Signaling, Danvers, MA, USA) and β-actin (loading control; Sigma). Bands were visualized using Carestream Image Station 4000R PRO with Carestream Molecular Imaging Software Version 5.0.2.30 (Carestream Health, Inc., New Haven, CT, USA) and quantified by UN-SCAN-IT Graph Digitizer Software 7.1 (Silk Scientific, Orem, UT, USA). The values were normalized to loading control and the normalized values for vehicle-treated and/or control-transfected cells were set to 1 for comparison within each cell line.
Muluhngwi P., Krishna A., Vittitow S.L., Napier J.T., Richardson K.M., Ellis M., Mott J.L, & Klinge C.M. (2016). Tamoxifen differentially regulates miR-29b-1 and miR-29a expression depending on endocrine-sensitivity in breast cancer cells. Cancer letters, 388, 230-238.
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3

Western Blot Analysis of DICER1 and DROSHA

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Western Blot analyses were performed as previously described [31 (link),34 (link)] with endothelial cells lysed using commercial lysis buffer (Cell Signaling Technology, Danvers, MA, USA) supplemented with a premade protease- and phosphatase-cocktail (Thermo Fisher Scientific). Cell debris were removed by centrifugation for 5 min at 15,000× g, 4 °C. SDS-PAGE was performed with equal amounts of protein per lane, followed by transfer to nitrocellulose membrane. Then, membranes were incubated with primary antibody solutions of DICER1 (Cell Signaling Technology; clone D38E7; 1:1000), DROSHA (Cell Signaling Technology; clone D28B1; 1:1000) and β-Actin (clone AC-15, Sigma Aldrich) according to the manufacturer’s instructions. Protein detection was achieved with peroxidase-conjugated secondary antibodies and the ECL system (Amersham Bioscience, Amersham, UK).
Braun H., Hauke M., Ripperger A., Ihling C., Fuszard M., Eckenstaler R, & Benndorf R.A. (2021). Impact of DICER1 and DROSHA on the Angiogenic Capacity of Human Endothelial Cells. International Journal of Molecular Sciences, 22(18), 9855.
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4

Quantitative Protein Expression Analysis

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Protein (30 μg) was subjected to 12% sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and electrophoretically transferred to a polyvinylidene fluoride membrane (Millipore, Billerica, MA). Protein blots were incubated separately with antibodies against Dicer1 (No. 3363, 1:500, Cell Signaling Technology, Beverly, MA), FABP1 (HPA028275, 1:100, Sigma-Aldrich, St. Louis, MO), or β-actin (1:4000, Sigma-Aldrich), probed with the appropriate alkaline phosphatase-conjugated secondary antibody, and detected using chemiluminescence. Immunoreactive protein bands were visualized by adding CDP-Star reagents (Roche Diagnostics, GmbH). Band signal intensities were quantified using Quantity One densitometric software (Bio-Rad, Hercules, CA).
Wu Y.L., Zhu Y.B., Huang R.D., Peng X.E, & Lin X. (2017). Multiple MicroRNAs Ameliorate Hepatocyte Steatosis and Injury by Suppressing FABP1 Expression. Cellular physiology and biochemistry : international journal of experimental cellular physiology, biochemistry, and pharmacology, 44(6).
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