circ_0001018 shRNA, circ_0001018 overexpression vectors, SOX4 overexpression vectors, miR-338-3p mimic, and miR-338-3p inhibitor were purchased from GenePharma (Shanghai, China). For the construction of the circ_0001018 overexpression and SOX4 overexpression vectors, the full length of circ_0001018 and SOX4 was amplified by PCR, and the primers containing BamHI and XhoI sites were designed. Next, the full length of circ_0001018 and the full-length of SOX4 were inserted into pLCDH-cir (GenePharma, China) and pcDNA3.1 (GenePharma, China), respectively, at the BamHI and EcoRI sites. Finally, puromycin was used to select the transfected cells. As for circ_0001018 shRNA, two shRNAs for knockdown of circ_0001018 were designed, synthesized, and inserted between the BamHI and EcoRI sites of the pLV-CMV-puro-U6-shRNA lentiviral vector. Puromycin also used to screen out the transfected cells. The sequences of circ_0001018 shRNA (sh-1 and sh-2), circRNA-OE, miR-338-3p mimic, and miR-338-3p inhibitor are given in Table S2 . The cell transfection was performed using circ_0001018 shRNA (sh-1 and sh-2), circ_0001018 overexpression vectors, SOX4 overexpression vectors, miR-338-3p mimic, and miR-338-3p inhibitor. After transfection for 48 h using Lipofectamine 3000 (Invitrogen), the transfected cells were collected to assess the transfection efficiency by qRT-PCR.
Mir 338 3p inhibitor
Manufactured by GenePharma
Sourced in China
The MiR-338-3p inhibitor is a laboratory product designed to inhibit the microRNA molecule miR-338-3p. MicroRNAs are small, non-coding RNA molecules that play a role in regulating gene expression. The MiR-338-3p inhibitor provides a tool for researchers to study the function and mechanisms of this specific microRNA.
Automatically generated - may contain errors
Lab products found in correlation
Mir 338 3p mimic, by GenePharma (3 mentions)
Lipofectamine 2000, by Thermo Fisher Scientific (2 mentions)
Plcdh cir, by GenePharma (1 mentions)
Pcdna3, by GenePharma (1 mentions)
Lipofectamine 3000, by Thermo Fisher Scientific (1 mentions)
Mir nc inhibitor, by GenePharma (1 mentions)
Lipofectamine rnaimax reagent, by Thermo Fisher Scientific (1 mentions)
Sirna1, by GenePharma (1 mentions)
Sirna2, by GenePharma (1 mentions)
Pcdna3.1 nr2f1 as1, by GenePharma (1 mentions)
Sh ccnd1, by GenePharma (1 mentions)
Sh nr2f1 as1, by GenePharma (1 mentions)
Lipofactamine 2000, by Thermo Fisher Scientific (1 mentions)
Mnng hos, by National Collection of Authenticated Cell Cultures (1 mentions)
Dmem f12, by Thermo Fisher Scientific (1 mentions)
Hfob1, by National Collection of Authenticated Cell Cultures (1 mentions)
Hfob1, by Thermo Fisher Scientific (1 mentions)
Streptomycin, by Thermo Fisher Scientific (1 mentions)
Penicillin, by Thermo Fisher Scientific (1 mentions)
Mg 63, by National Collection of Authenticated Cell Cultures (1 mentions)
5 protocols using mir 338 3p inhibitor
1
Circular RNA Regulation in Cancer
2
LINC00707 Silencing and miR-338-3p Modulation
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Small interfering RNA (siRNA) targeting LINC00707 (siRNA1: 5′-GGCUUUCCAUGACCCAUAAUU-3′, siRNA2: 5′-GGAAGCCACU CCUGCAUUUUU-3′, siRNA3: 5′-GCAGGAACAUCACCAUCUUUU-3′), negative control siRNA (si-NC), miR-338-3p mimic, negative control miRNA (miR-NC), miR-338-3p inhibitor, and miR-NC inhibitor were synthesized using GenePharma Co., Ltd (Shanghai, China). Lipofectamine RNAiMAX Reagent (Invitrogen, Carlsbad, CA, USA) was used for cell transfection according to the manufacturer’s instructions. To analyze the effect of LINC00707 silencing, MG-63 or Saos-2 cells were transfected with siNC or siRNA1 and classified as the siNC group and the siRNA1 group. To analyze whether the miR-338-3p inhibitor weakens the effect of LINC00707 silencing, MG-63 or Saos-2 cells were transfected with siRNA1 plus miR-338-3p inhibitor group or siRNA1 plus miR-NC inhibitor and classified as the siRNA1 + miR-338-3p inhibitor group and the siRNA1 + miR-NC inhibitor group. After transfected at 24 h, LINC00707 expression was measured by quantitative reverse transcription PCR (qRT-PCR).
3
Silencing NR2F1-AS1 and CCND1 in Thyroid Cells
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ShRNA negative control (NC), sh‐NR2F1‐AS1, sh‐CCND1, miR‐338‐3p mimics, miR‐338‐3p inhibitor and NC were all provided by GenePharma Co, Ltd (Shanghai, China). In brief, sh‐NR2F1‐AS1 was synthesized and loaded into the pENTRTM/U6 vector. After finishing this construction, pENTRTM/U6‐sh‐NR2F1‐AS1 was sequenced to verify the accuracy of the inserted sequence. Thereafter, pENTRTM/U6‐sh‐NR2F1‐AS1 was introduced into FTC‐133 cells and B‐CPAP cells using Lipofactamine2000 (Invitrogen, Carlsbad, CA). Stably transfected cell with antibiotic resistance were sifted and enriched by adding puromycin to culture medium. Based on protocols of Lipofectamine 2000 (Invitrogen), miR‐338‐3p mimics, miR‐338‐3p inhibitor and NC were introduced into FTC‐133 cells and B‐CPAP cells. PcDNA3.1‐NR2F1‐AS1 and pcDNA3.1‐CCND1 were from GenePharma as well. Before plasmid transfection, FTC‐133 cells and B‐CPAP cells were suspended and seeded in six‐well culture plates at 37°C. When cell confluence rate reached 80% to 90%, cell culture media should be replaced by serum‐free fresh medium 3 hours before transfection.
4
Osteosarcoma Cell Line Culture Protocol
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Human osteosarcoma cell lines (MNNG/HOS, MG-63) and a normal osteoblast cell line (hFOB1.19) were purchased from the Cell Bank of Type Culture Collection of Chinese Academy of Sciences (Shanghai, China). The osteoblast cells hFOB1.19 was cultured in Dulbecco's modified Eagle's medium/ham's F‐12 (DMEM/F‐12; Gibco, Grand Island, NY) containing 10% fetal bovine serum (FBS; Gibco) and 0.3 mg/ml G418. The MNNG/HOS and MG-63 cell lines were grown in Eagle's Minimum Essential Medium (MEM; Gibco) supplemented with 10% FBS, 100‐units/ml penicillin, and 100 mg/ml streptomycin (Gibco). The cells were incubated at 37°C in 5% CO2 incubator.
The pcDNA (Vector) as well as pcDNA-NRCAM-1 overexpression (NRCAM-OE) plasmids were provided by GenePharma (Shanghai, China). The pcDNA (Vector) and pcDNA-KLF9 overexpression (KLF9-OE) plasmids were obtained from GenePharma (Shanghai, China). The miR-338-3p mimic, miR-338-3p inhibitor and the mimics negative control (miR-NC) were purchased from GenePharma (Shanghai, China). All transfections were executed by applying Lipofectamine 3000 reagent (Invitrogen, USA) in accordance to the manufacturer's protocols. Then cells were cultured for 48 hours following by collection for the validation of transfection efficiency via quantitative reverse transcription-polymerase chain reaction (qRT-PCR) analysis.
The pcDNA (Vector) as well as pcDNA-NRCAM-1 overexpression (NRCAM-OE) plasmids were provided by GenePharma (Shanghai, China). The pcDNA (Vector) and pcDNA-KLF9 overexpression (KLF9-OE) plasmids were obtained from GenePharma (Shanghai, China). The miR-338-3p mimic, miR-338-3p inhibitor and the mimics negative control (miR-NC) were purchased from GenePharma (Shanghai, China). All transfections were executed by applying Lipofectamine 3000 reagent (Invitrogen, USA) in accordance to the manufacturer's protocols. Then cells were cultured for 48 hours following by collection for the validation of transfection efficiency via quantitative reverse transcription-polymerase chain reaction (qRT-PCR) analysis.
5
Modulating miR-338-3p and MMP-2 Expression
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The overexpression and knockdown vectors of miR-338-3p and MMP-2 were purchased from Shanghai GenePharma (Shanghai, China) named as miR-338-3p mimics, miR-338-3p inhibitor, pc-MMP-2, and si-MMP-2, which were in comparison with negative control (NC) named as NC mimic, NC inhibitor, pc-NC, and si-NC, respectively. When the cells’ culture confluency reached 50%, the overexpression and knockdown vectors of miR-338-3p, MMP-2, and NC were transfected into the cells with Lipofectamine-2000 (Invitrogen, Carlsbad, CA, USA) following the protocol of the manufacturer. Transfected cells were harvested for 24, 48, or 72 h for further experiments.
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