Mimics and inhibitors of miR-17-5p or miR-20a, cholesterol-conjugated miR-17-5p and miR-20a inhibitors, c-myc, uPAR, and DR5-specific small interfering RNA (siRNA) were chemically synthesized by RiboBio Co., Ltd. (Guangzhou, China). The sequence of siRNAs for c-myc, uPAR, and DR5 were as follows: c-myc, 5′-CTATGACCTCGACTACGAC-3′; uPAR, 5′-GCTGTACCCACTCAGAGAA-3′; DR4 5′-CUCUGAUGCUGUUCUUUGAtt and DR5, 5′-AAGACCCUUGUGCUCGUUGUC-3′. The following reagents and antibodies were obtained as indicated: DR5 antibody was from Santa Cruz Biotechnology (Dallas, TX,, USA); procaspase 8, caspase 3/8, and total and phosphorylated forms of ERK antibodies were from Cell Signaling Biotech; uPAR antibody was from R&D Systems (Minneapolis, MN, USA); and c-myc, actin, GAPDH, and horseradish peroxidase (HRP)-conjugated secondary antibodies were from Zhongshan Goldenbridge Biotechnology (Beijing, China). The ECL-Plus chemiluminescence system was from Applygen Technologies (Beijing, China).
C myc
Manufactured by RiboBio
Sourced in China
C-myc is a lab equipment product used for measuring the expression levels of the c-myc gene. It provides a reliable and quantitative method for analyzing the transcriptional activity of this important oncogene.
Automatically generated - may contain errors
Lab products found in correlation
Lipofectamine 2000, by Thermo Fisher Scientific (3 mentions)
Ecl plus chemiluminescence system, by Applygen (1 mentions)
C myc, by Zhongshan Golden Bridge Biotechnology (1 mentions)
C myc, by Santa Cruz Biotechnology (1 mentions)
Horseradish peroxidase hrp conjugated secondary antibody, by Zhongshan Golden Bridge Biotechnology (1 mentions)
Gapdh, by Zhongshan Golden Bridge Biotechnology (1 mentions)
Tead1, by RiboBio (1 mentions)
Sic myc, by RiboBio (1 mentions)
Transfection reagent, by Thermo Fisher Scientific (1 mentions)
Puromycin, by Merck Group (1 mentions)
5 protocols using c myc
1
Inhibition of miR-17-5p and miR-20a
2
Transcriptional Regulation by GCN5, E2F1, and c-Myc
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Cells were seeded in 6‐cm dishes at 3 × 104 cells and cultured in medium without antibiotics for 24 hour. Chemically modified Stealth small interfering RNA (siRNA) targeting GCN5, E2F1, c‐Myc and control siRNA were purchased from Guangzhou RiboBio (RiboBio, Guangzhou, China) and transfected into cells using Lipofectamine 2000 (Invitrogen, Life Technologies, CA, USA) according to manufacturer's instructions. Cells were transfected with siRNA at a concentration of 20 nmol/L. The sequences of siRNAs are as follows: siGCN5‐1 5′‐GGAAAUGCAUCCUGCAGAU‐3′; siGCN5‐2 5′‐GAGGCCUCAUUGACAAGUA‐3′; siE2F1‐1 5′‐GUCACGCUAUGAGACCUCA‐3′; siE2F1‐2 5′‐GGACCUUCGUAGCAUUGCA‐3′; sic‐Myc‐1 5′‐CGUCCAAGCAGAGGAGCAA‐3′; sic‐Myc‐2 5′‐CCAAGGUAGUUAUCCUUAA‐3′.
Thirty‐six hours after transfection, the cells were treated with DMSO or 10 μg/mL bleomycin and incubated for an additional 36 hours. Cells were harvested for protein knockdown analysis by Western blotting or for cell cycle analysis by flow cytometry.
Thirty‐six hours after transfection, the cells were treated with DMSO or 10 μg/mL bleomycin and incubated for an additional 36 hours. Cells were harvested for protein knockdown analysis by Western blotting or for cell cycle analysis by flow cytometry.
3
Targeted Gene Knockdown and Knockout in Cells
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siRNAs targeting PHF8, JARID1A, JARID1C, JHDM1A, JHDMA2A, UTX, PHF2, TEAD1, GLUL, FASN, and c-MYC were purchased from RiboBio (RiboBio Co. Ltd., Guangzhou, Guangdong, China). siRNAs targeting TES, FRG1, GRIPAP1, LIN28A, and N6AMT1 were synthesized by Sangon (Sangon Biotech, Shanghai, China). All siRNAs were used to transiently knock down the indicated genes, with scrambled siRNA as the control. In brief, cells were cultured until 70 to 80% confluence in six-well were infected with 50 nM siRNAs in presence of 5 μl of Lipofectamine 2000 (11668019, Thermo Fisher Scientific) according to the manufacturer’s protocol. After 24 hours of transfection, the cells were harvested for next manipulation. To stably knock out PHF8, sgRNA (four sgRNAs targeting human PHF8) was cloned into lentiCRISPR v2 vector and then expressed in lentivirus. Cells with 70 to 80% confluence were infected by lentivirus containing sgRNAs targeting PHF8 in presence of polybrene (8 ng/ml), and scrambled sgRNA was used as control. After 48 hours of transfection, puromycin was added to select infected cells until 5 days. Then, a monoclonal cell population was isolated by limiting dilution method. The siRNA and sgRNA sequences are listed in table S8.
4
Targeted Silencing of SNHG17, c-Myc, and LRPPRC
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The small interfering RNAs (siRNAs) targeting the transcripts of human SNHG17 (GeneBank accession No. NR_152762.1), c-Myc (NM_002467.6) and LRPPRC (NM_133259.4) were designated as siSNHG17, sic-Myc, and siLRPPRC, respectively, and were purchased from Ribobio (Guangzhou, China). The negative control (NC) RNA duplex for siRNA was non-homologous to any human genome sequences.
5
Targeted Silencing of URGCP, NF-κB/p65, and c-myc
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siRNAs targeting URGCP, NF-κB/p65 and c-myc were purchased from RIBOBIO. A nonsilencing siRNA oligonucleotide that didn’t target any mammalian gene was used as a negative control. Transfection of siRNA duplexes was performed using the transfection reagent (Invitrogen) according to the manufacturer’s instruction. shURGCP Lentiviral particle was purchased from GENECHEM (CHN), and a control shRNA was used as control. U87 and U251 were transfected with shURGCP and selected with puromycin (Sigma).
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