The animal experiment was approved by the Animal Ethics Committee of Huaihe Hospital of Henan University and performed in accordance with the guidelines of the National Animal Care and Ethics Institution. All animal procedures were conducted at the Animal Institute of Henan University. 143B cells were transfected with lentivirus containing circ_0056285 short hairpin RNA (sh-circ) or the control (sh-NC) (GenePharma, Shanghai, China). The stable transfected 143B cells (5×106) were subcutaneously injected into the right-back of BALB/c nude mice. The tumor size was measured using a digital caliper at indicated time points. After 35 days, the mice were sacrificed by cervical dislocation. If no spontaneous breathing was observed within 2–3 min and no blink reflex, the mice were considered euthanized. Then, the tumors were excised and weighed.
Sh circ
Manufactured by GenePharma
Sourced in China
Sh-circ is a laboratory equipment designed for the production of short hairpin circular RNA (sh-circ). It provides a reliable and efficient platform for the generation of circular RNA molecules, which can be used in various research applications.
Automatically generated - may contain errors
Lab products found in correlation
Sh nc, by GenePharma (2 mentions)
Lipofectamine 3000, by Thermo Fisher Scientific (2 mentions)
Sw480, by Health Science Research Resources Bank (1 mentions)
Caco2, by Health Science Research Resources Bank (1 mentions)
Mir 149 5p mimic, by GenePharma (1 mentions)
Lipofectamine 2000, by Thermo Fisher Scientific (1 mentions)
Mir nc, by GenePharma (1 mentions)
Fetal bovine serum (fbs), by Procell (1 mentions)
Si col1a1, by GenePharma (1 mentions)
Sw1353, by Procell (1 mentions)
Si nc, by GenePharma (1 mentions)
Penicillin streptomycin, by Procell (1 mentions)
Saos 2, by Procell (1 mentions)
Hfob1, by Procell (1 mentions)
5 protocols using sh circ
1
Investigating Circular RNA in Osteosarcoma
2
Colon Cancer Cell Line Manipulation
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Human colon cancer cell lines (CACO2, HT29, LOVO, SW620, and SW480) and normal colon cell line NCM460 were purchased from Health Science Research Resources Bank. Cells were cultured in Dulbecco’s modified Eagle medium supplemented with 10% fetal bovine serum (FBS) at 37% and 5% CO2. ShRNA against negative control (sh-NC), shRNA against circCTNNA1 (sh-circ), pcDNA-based circCTNNA1 overexpression vector (circCTNNA1), pcDNA vector, miR-149-5p mimic, negative control mimic (NC mimic), miR-149-5p inhibitor, and negative control inhibitor (NC inhibitor) were synthesized by GenePharma (Shanghai, China) and transfected with Lipofectamine 3000 (Invitrogen, USA).
3
Silencing circCAMSAP1 expression
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The recombinant lentivirus vector sh-circ (to silence circCAMSAP1 expression) and NC (negative control) were constructed and packaged by Gene Pharma (Shanghai, China). The miR-1182 mimic and miR-NC (negative control) were purchased from Solarbio (Beijing, China). pcDNA3.1‐BIRC5 (a vector lacking the BIRC5 3ʹ-UTR) was obtained from Gene Pharma. The transfection assay was performed using Lipofectamine 2000 (Invitrogen, Carlsbad, CA, USA).
4
Knockdown of circ_0030235 in H9c2 cells
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Short hairpin (sh) RNA targeting circ_0030235 (sh-circ) and its corresponding control (sh-NC) was synthesized and inserted into vectors by GenePharma (Shanghai, China). MiR-526b inhibitor and its corresponding control (miR-NC) were purchased from GenePharma. A transient transfection was performed using Lipofectamine 3000 (Thermo Fisher Scientific, Waltham, MA, USA). In brief, 48 h prior to transfection, 5 × 105 H9c2 cells were plated in a 6-well plate. Cells were transfected with 1 µg shRNA vectors or 0.1 µmol miR inhibitor. After 48 h, cells were subjected to total RNA extraction for qRT-PCR detection.
5
Osteoblast and Osteosarcoma Cell Culture
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Normal osteoblasts (hFOB1.19) and osteosarcoma cells (HOS, Saos-2, and SW1353) were obtained from ProCell (China). SW1353 (Leibovitz’s L-15 Medium; Procell), Saos-2 (McCoy’s 5A; Procell), HOS (MEM; Procell), and hFOB1.19 (DMEM/F12; Procell) were maintained in media containing 1% penicillin-streptomycin (Procell) and 10% fetal bovine serum (FBS, Procell). Culture was performed under a 5% CO2 atmosphere at 37°C.
Small interfering RNAs (siRNAs) targeting circ_0020378 (si-circ_0020378), COL1A1 (si-COL1A1), miR-339-3p inhibitor and mimic, and their respective negative controls (si-NC, inhibitor NC, and mimic NC) were purchased from GenePharma, China. Recombinant lentiviruses (multiplicity of infection [MOI] = 50) carrying short hairpin RNAs (shRNAs) targeting either circ_0020378 (sh-circ) or sh-NC were purchased from GenePharma. Lipo3000 was used to introduce synthetic nucleotides (siRNAs, mimics, and inhibitors) into Saos-2 and HOS cells. After a 48-h transfection period, the efficacy of transfection was verified by RT-qPCR. For shRNA lentivirus infection, approximately 80% confluent Saos-2 cells were incubated with the virus particles (MOI = 50). After 14 days, the survivors were screened by adding 0.5 μg/mL puromycin. Positive monoclonal cells were expanded and verified using RT-qPCR.
Small interfering RNAs (siRNAs) targeting circ_0020378 (si-circ_0020378), COL1A1 (si-COL1A1), miR-339-3p inhibitor and mimic, and their respective negative controls (si-NC, inhibitor NC, and mimic NC) were purchased from GenePharma, China. Recombinant lentiviruses (multiplicity of infection [MOI] = 50) carrying short hairpin RNAs (shRNAs) targeting either circ_0020378 (sh-circ) or sh-NC were purchased from GenePharma. Lipo3000 was used to introduce synthetic nucleotides (siRNAs, mimics, and inhibitors) into Saos-2 and HOS cells. After a 48-h transfection period, the efficacy of transfection was verified by RT-qPCR. For shRNA lentivirus infection, approximately 80% confluent Saos-2 cells were incubated with the virus particles (MOI = 50). After 14 days, the survivors were screened by adding 0.5 μg/mL puromycin. Positive monoclonal cells were expanded and verified using RT-qPCR.
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