The miRNA-506 mimic and the negative control were obtained from GenePharma (Shanghai, China). The sequence of the miR-506 mimic was 5′-UAAGGCACCCUUCUGAGUAGA-3′. Cells (5 × 105 cells/2 ml/well) were seeded at 60% confluence in a six-well plate. After 48 h, the miRNA-506 mimic or the negative control was transfected into cells using Lipofectamine 2000 (Invitrogen, USA) at a final concentration of 50 nM according to the manufacturer's instructions. The siRNA targeting EZH2, with a sequence of 5′-AAGACTCTGAATGCAGTTGCT-3′, and the miR-506 inhibitor was purchased from GenePharma. The transfection of siRNA and miR-506 inhibitor was performed as described above. The final siRNA and miR-506 inhibitor concentration were 100 nM and 50 nM.
Mir 506 inhibitor
Manufactured by GenePharma
Sourced in China
The MiR-506 inhibitor is a lab equipment product designed to inhibit the expression of the miR-506 microRNA. It is intended for use in research applications to study the biological functions and regulatory mechanisms of miR-506.
Automatically generated - may contain errors
Lab products found in correlation
Lipofectamine 2000, by Thermo Fisher Scientific (2 mentions)
Lipofectamine 3000, by Thermo Fisher Scientific (1 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (1 mentions)
Pcdna3.1 vector, by Thermo Fisher Scientific (1 mentions)
Mir nc, by GenePharma (1 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions)
Inh nc, by GenePharma (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
3 protocols using mir 506 inhibitor
1
Investigating miR-506 Regulation of EZH2
2
Targeted Silencing of NEAT1 and LIN28B
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The NEAT1-shRNA-Lentivirus for use in the in vivo experiments was purchased from the ABM Company. LIN28B-related stable cell lines were constructed according to previously published guidelines51 (link). siRNAs targeting NEAT1 and LIN28B were purchased from Suzhou Synbio Technologies and transfected into cells using Lipofectamine 3000 (Invitrogen). All siRNA and shRNA sequences are provided in Supplementary Table 5 . miR-506 inhibitor was purchased from GenePharma (Shanghai, China).
3
Glioma Cell Line Manipulation and Characterization
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Human glioma cell lines (U87, U251, SHG44, A172) were purchased from the Institute of Biochemistry and Cell Biology of the Chinese Academy of Sciences (Shanghai, China), as well as normal human astrocytes (NHAs). Cells were cultured with RPMI-1640 medium (Invitrogen, Carlsbad, CA, USA) supplemented with 10% fetal bovine serum (FBS, Invitrogen) and 100 U/mL penicillin/streptomycin (Invitrogen) in a humidified incubator with 5% CO2 at 37 °C.
To construct LINC00963-overexpressing cells, the full-length human LINC00963 sequence was amplified by PCR, and the product was subcloned into a pcDNA3.1 vector (Invitrogen) and named pcLINC00963. Specific siRNAs were designed to knockdown LINC00963 (si-RNA1, 5ʹ-GGTTCCTCATCTGCCAGTT-3ʹ; si-RNA2, 5ʹ-GGCGCAGTAACAATATAAT-3ʹ), and si-NC was used as negative control. A negative control (pcNC) was also constructed. MiR-506 mimic, miR-506 inhibitor and their corresponding negative controls (miR-NC and inh-NC) were purchased from GenePharma (Shanghai, China). BCAT1-expressing vectors (named pcBCAT1) were generated by GenePharma and used to restore the expression of BCAT1 in cells, and pcDNA was used as a control. The transfection procedure was performed using Lipofectamine 2000 (Invitrogen) according to the manufacturer’s instructions.
To construct LINC00963-overexpressing cells, the full-length human LINC00963 sequence was amplified by PCR, and the product was subcloned into a pcDNA3.1 vector (Invitrogen) and named pcLINC00963. Specific siRNAs were designed to knockdown LINC00963 (si-RNA1, 5ʹ-GGTTCCTCATCTGCCAGTT-3ʹ; si-RNA2, 5ʹ-GGCGCAGTAACAATATAAT-3ʹ), and si-NC was used as negative control. A negative control (pcNC) was also constructed. MiR-506 mimic, miR-506 inhibitor and their corresponding negative controls (miR-NC and inh-NC) were purchased from GenePharma (Shanghai, China). BCAT1-expressing vectors (named pcBCAT1) were generated by GenePharma and used to restore the expression of BCAT1 in cells, and pcDNA was used as a control. The transfection procedure was performed using Lipofectamine 2000 (Invitrogen) according to the manufacturer’s instructions.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!