All animal procedures were approved by the Alfred Medical Research and Education Precinct (AMREP) Animal Ethics Committee in accordance with the National Health and Medical Research Council of Australia guidelines. The animals were maintained on a 12-hour light/dark cycle with unrestricted access to water and standard chow (Specialty Feeds, Glen Forrest, WA, Australia) under pathogen-free conditions in the Precinct Animal Centre of the Baker IDI Heart and Diabetes Institute. Nox4−/− [21 (link)] mice were backcrossed with Apoe−/− mice on the C57BL/6J background (B6.129P2-Apoetm1Unc /J, Jax Labs, Bar Harbor, ME, USA) for 10 generations to generate Nox4−/−Apoe−/− double knockout animals (Transnetyx, USA); as previously described in [9 ]. Diabetes was induced in six-week-old Nox4−/−Apoe−/− and Nox4−/−Apoe−/− wildtype male mice by five daily i.p. injections of streptozotocin [55 mg/kg dissolved in citrate buffer (Sigma-Aldrich, St Louis, MO, USA)] [22 (link)]. Mice with a blood glucose level ≥15 mmol/L were deemed diabetic and included in the study. After 20 weeks, the mice were killed by an i.p. injection of sodium pentobarbital (100 mg/kg; Euthatal, Sigma-Aldrich, Castle Hill, NSW, Australia).
Standard chow
Manufactured by Specialty Feeds
Sourced in Australia
Standard chow is a laboratory animal diet formulated to provide a complete and balanced nutrition for rodents and other small laboratory animals. It contains a mixture of grains, proteins, vitamins, and minerals essential for the animals' health and growth.
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Lab products found in correlation
4 protocols using standard chow
1
Diabetic Atherosclerosis in Nox4 Knockout Mice
2
Ethical Animal Research Protocol
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All animal work was performed at the Baker Heart and Diabetes Institute with appropriate ethics approval (approved by the Alfred Medical Research and Education Precinct Animal Ethics Committee, project number E/1793/2018/B) and in accordance with the Australia Code for the Care and Use of Animals for Scientific Purposes, 8th Edition (2013). All animal breeding, transportation and experimentation was conducted in accordance with the Commonwealth of Australia Gene Technology Act 2000 and Gene Technology Regulations 2001 and approved by the relevant Institutional Biosafety Committees (notifiable low risk dealings N80, N143 and 11974). Experimental procedures were performed in accordance with locally approved safe work procedures and risk assessments. Mice were housed on a 12-h light cycle (lights on 0100 h), with standard chow (Specialty Feeds, Australia; 19% protein, 5% fat, 5% fiber, 0.2% sodium) and water available ad libitum (except for temporary fasting prior to feeding behavioral tests). Mice were group-housed prior to radiotelemeter implantation, then individually for the remaining experimental program.
3
ApoE-Deficient Mice Model Treated with Tan IIA
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All animal procedures were approved by the Ethics Committee of Henan Province People's Hospital. ApoE−/− mice with a C57BL/6J background were purchased from Beijing Biocytogen (Beijing, China). All animals were kept on a regular dark/light cycle, with unrestricted access to water and standard chow (Specialty Feeds, Glen Forrest, WA, Australia) under pathogen-free conditions.
Sixteen male ApoE−/− mice at the age of 6 weeks were randomly divided into two groups of eight animals each: control group (NC) and Tan IIA (30 mg/kg) group. Mice in Tan IIA groups were gavaged with Tan IIA (30 mg/kg) suspended in 0.5% sodium carboxymethyl cellulose (CMC-Na) daily for 20 weeks, whereas mice in control groups were gavaged with 0.5% CMC-Na. After 20 weeks, the mice were killed by an i.p. injection of sodium pentobarbital (100 mg/kg; Euthatal, Sigma-Aldrich, Castle Hill, NSW, Australia).
Sixteen male ApoE−/− mice at the age of 6 weeks were randomly divided into two groups of eight animals each: control group (NC) and Tan IIA (30 mg/kg) group. Mice in Tan IIA groups were gavaged with Tan IIA (30 mg/kg) suspended in 0.5% sodium carboxymethyl cellulose (CMC-Na) daily for 20 weeks, whereas mice in control groups were gavaged with 0.5% CMC-Na. After 20 weeks, the mice were killed by an i.p. injection of sodium pentobarbital (100 mg/kg; Euthatal, Sigma-Aldrich, Castle Hill, NSW, Australia).
4
Characterization of Vglut1-Cre Mouse Model
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All experiments were conducted in accordance with the Monash Animal Ethics Committee guidelines. C57BL/6 male mice were obtained from the Monash Animal Services facility. Vglut1-ires-cre male mice on a C57BL/6 background were obtained from the Jackson Laboratory (B6.Slc17a7-IRES2-Cre-D; stock number 023527) and bred in the Monash Animal Services Facility. All mice were aged between 8-10 weeks at beginning of experiments. Mice were maintained on a 12-hour light-dark cycle with ad libitum access to standard chow (Specialty Feeds, Western Australia) and water under standard laboratory conditions (21°C).
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