THJ-11T (KRAS, TP53, TERT), THJ-16T (PI3KCA, TP53, TERT), THJ-21T (BRAF, TP53, TERT) and THJ-29T (APC, TP53, TERT) ATC cell lines were originated in our laboratory (Marlow et al. 2010 (link)) and were short-tandem repeat (STR) verified and validated to the respective patient’s ATC tissue. The APC mutation for THJ-29T was identified by Drs. James Fagin and Jeffrey Knauf as well as validating the other mutations (personal communication). Thyroid cells were maintained in RPMI 1640 medium (Cellgro, Manassas, VA) supplemented with 5% fetal bovine serum (Hyclone, Logan, UT), non–essential amino acids (Cellgro), sodium pyruvate (Cellgro), HEPES (Cellgro) and penicillin-streptomycin-amphotericin B (Cellgro) at 37°C in a humidified atmosphere with 5% CO2. 293FT cells were purchased from (Invitrogen, Carlsbad, CA) and were maintained in DMEM as per the manufacturer’s protocol along with 500 μg/ml neomycin (MP Biomedical, Solon, OH).
Neomycin
Manufactured by MP Biomedicals
Sourced in United States
Neomycin is an aminoglycoside antibiotic used in laboratory settings. It is effective against a variety of gram-positive and gram-negative bacteria.
Automatically generated - may contain errors
Lab products found in correlation
Vancomycin, by MP Biomedicals (5 mentions)
Ampicillin, by MP Biomedicals (5 mentions)
Vancomycin, by Avantor (2 mentions)
Metronidazole, by Thermo Fisher Scientific (2 mentions)
Qiaamp fast dna stool mini kit, by Qiagen (2 mentions)
Ampicillin, by Merck Group (2 mentions)
Gentamicin, by Merck Group (2 mentions)
Sodium pyruvate, by Corning (1 mentions)
Nonessential amino acid, by Corning (1 mentions)
Hepes, by Corning (1 mentions)
Rpmi 1640 medium, by Corning (1 mentions)
Penicillin streptomycin amphotericin b, by Corning (1 mentions)
293ft cell, by Thermo Fisher Scientific (1 mentions)
Gentamycin, by MP Biomedicals (1 mentions)
Erythromycin, by MP Biomedicals (1 mentions)
9 protocols using neomycin
1
Anaplastic Thyroid Cancer Cell Lines
2
Chronic Unpredictable Mild Stress in Mice
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Mice were subjected to CUMS for a total duration of 6 weeks (Figure 1 ). With the exception of those in the control group, all animals were subjected to the mild stress protocol in an unpredictable manner for 6 weeks. The protocol consisted of seven stressors: food deprivation for 24 h, water deprivation for 24 h, restraint stress for 5 h, overnight illumination for 8 h, horizontal oscillation for 20 min, cage tilting at 45° for 24 h, and a soiled cage environment (500 mL water added to 250 g sawdust bedding) for 24 h. Antibiotic treatment was provided as previously described 20 (link). Briefly, animals were treated with gentamycin (100 mg/L; MPbio), ampicillin (1 g/L; MPbio), erythromycin (10 mg/L; MPbio), vancomycin (0.5 g/L; MPbio), and neomycin (0.5 g/L; MPbio), which were administered via drinking water for 6 weeks. Mice were then divided into six groups for analysis (WT, HZ, WT+CUMS, HZ+CUMS, WT+CUMS+antibiotics, HZ+CUMS+antibiotics).
3
Synthesis of Neomycin Compound
4
Gut Microbiome Alterations in Mettl3 Mice
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Mettl3flox/flox and Mettl3VilCreERΔ/Δ mice were moved to new cages at 7 weeks of age and given sterile DI water supplemented with 0.5g/L Ampicillin (Sigma A9518), 0.5g/L Neomycin (MP Biomedicals 180610), 0.5g/L Gentamicin (Sigma G1914–5G), 0.25g/L Vancomycin (VWR 0990), 0.25g/L Metronidazole (Thermo 210340050), and 4g/L Splenda to enhance taste. After seven days, mice were injected with tamoxifen for four days as described above and stool collected on the final day of injection. Stool from Mettl3flox/flox and Mettl3VilCreERΔ/Δ mice on normal drinking water was used as controls. DNA was extracted from stool using the QIAamp Fast DNA Stool Mini Kit (Qiagen 51604) and quantitative PCR was performed as described in the qPCR methods section using primer Ba04230899_s1. qPCR quantification was normalized to stool weight.
5
Gut Microbiome Modulation Protocol
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Mettl3fl/fl and Mettl3VilCreERΔ/Δ mice were moved to new cages at 7 weeks of age and given sterile deionized water supplemented with 0.5 g/L ampicillin (MilliporeSigma A9518), 0.5 g/L neomycin (MP Biomedicals 180610), 0.5 g/L gentamicin (MilliporeSigma G1914-5G), 0.25 g/L vancomycin (VWR 0990), 0.25 g/L metronidazole (Thermo Fisher Scientific 210340050), and 4 g/L Splenda to enhance taste. After 7 days, mice were injected with tamoxifen for 4 days as described above, and stool was collected on the final day of injection. Stool from Mettl3fl/fl and Mettl3VilCreERΔ/Δ mice on normal drinking water was used as controls. DNA was extracted from stool using the QIAamp Fast DNA Stool Mini Kit (QIAGEN 51604), and qPCR was performed as described in the PCR Methods section using primer Ba04230899_s1. qPCR quantification was normalized to stool weight.
6
Antibiotic Treatment in Mice
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Vancomycin (50 mg/kg, CAS: 123409-00-7, MP bio, California, USA), neomycin (100 mg/kg, CAS: 1405-10-3, MP bio), and metronidazole (100 mg/kg, CAS: 443-48-1, MCE, New Jersey, USA) were mixed using sterile drinking water [4 (link)]. Then the mixture was orally gavaged to 3-week-old or 6-week-old SPF C57BL/6J mice twice a day for five or seven consecutive days and the amount of infusion was based on the weight of mice. The mixture was prepared every day and used freshly. During the treatment, ampicillin (1 mg/mL, CAS: 69-52-3, MP bio) was added into the drinking water of mice and changed with fresh solution every 3 days. For 3-week-old SPF C57BL/6J mice, the antibiotic treatment lasted for 5 days [53 (link)]. All the mice were handled aseptically.
7
Antibiotic Treatment in Young Mice
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Vancomycin (50 mg/kg, CAS: 123409-00-7, MP bio, California, USA), neomycin (100 mg/kg, CAS: 1405-10-3, MP bio) and metronidazole (100 mg/kg, CAS: 443-48-1, MCE, New Jersey, USA) were mixed using sterile drinking water [4] . Then the mixture was orally gavaged to 3-week-old or 6-week-old SPF C57BL/6J mice twice a day for 5 or 7 consecutive days and the amount of infusion was based on the weight of mice. The mixture was prepared every day and used freshly. During the treatment, ampicillin (1 mg/mL, CAS: 69-52-3, MP bio) was added into the drinking water of mice and changed with fresh solution every 3 days. For 3-week-old SPF C57BL/6J mice, the antibiotic treatment lasted for 5 days [53] . All the mice were handled aseptically.
8
Antibiotic Cocktail for Mouse Gut Microbiome
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Vancomycin (50 mg/kg, CAS: 123409-00-7, MP bio, California, USA), neomycin (100 mg/kg, CAS: 1405-10-3, MP bio) and metronidazole (100 mg/kg, CAS: 443-48-1, MCE, New Jersey, USA) were mixed using sterile drinking water [4] . Then the mixture was orally gavaged to 3-week-old or 6-week-old SPF C57BL/6J mice twice a day for 5 or 7 consecutive days and the amount of infusion was based on the weight of mice. The mixture was prepared every day and used freshly. During the treatment, ampicillin (1 mg/mL, CAS: 69-52-3, MP bio) was added into the drinking water of mice and changed with fresh solution every 3 days. For 3-week-old SPF C57BL/6J mice, the antibiotics treatment lasted for 5 days [53] . All the mice were handled aseptically.
9
Antibiotic Cocktail for Mouse Gut Microbiome
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+ Expand
Vancomycin (50 mg/kg, CAS: 123409-00-7, MP bio, California, USA), neomycin (100 mg/kg, CAS: 1405-10-3, MP bio) and metronidazole (100 mg/kg, CAS: 443-48-1, MCE, New Jersey, USA) were mixed using sterile drinking water [4] . Then the mixture was orally gavaged to 3-week-old or 6-week-old SPF C57BL/6J mice twice a day for 5 or 7 consecutive days and the amount of infusion was based on the weight of mice. The mixture was prepared every day and used freshly. During the treatment, ampicillin (1 mg/mL, CAS: 69-52-3, MP bio) was added into the drinking water of mice and changed with fresh solution every 3 days. For 3-week-old SPF C57BL/6J mice, the antibiotics treatment lasted for 5 days [53] . All the mice were handled aseptically.
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