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C57bl 6j b6 male mice

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C57BL/6J (B6) male mice are a commonly used inbred mouse strain. They are widely utilized in biomedical research due to their well-characterized genetic background and various physiological traits.

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Lab products found in correlation

Penicillin, by Thermo Fisher Scientific (2 mentions) Streptomycin, by Thermo Fisher Scientific (2 mentions) B27 supplement, by Thermo Fisher Scientific (2 mentions) Ham s f12 medium, by Merck Group (2 mentions) Glutamax, by Thermo Fisher Scientific (2 mentions) Purelink rna mini kit, by Thermo Fisher Scientific (2 mentions) Insulin, by Merck Group (2 mentions) Epidermal growth factor (egf), by Merck Group (2 mentions) Sodium pyruvate, by Corning (2 mentions) Penicillin, by American Type Culture Collection (2 mentions) Streptomycin, by American Type Culture Collection (2 mentions) Nih3t3 cells, by American Type Culture Collection (2 mentions) Mouse ref 8 whole genome microarrays, by Illumina (2 mentions) Nor ltj, by Jackson ImmunoResearch (1 mentions) Nod shiltj mice, by Jackson ImmunoResearch (1 mentions) Prolab rmh 180 5ll2 chow, by TestDiet (1 mentions)

8 protocols using c57bl 6j b6 male mice

1

Generation and Analysis of Transgenic Mice

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C57BL/6J (B6) male mice, female non-obese diabetic (NOD/ShiLtJ) mice, female non-obese diabetes-resistant (NOR/LtJ), and INS1cre (B6(Cg)-Ins1tm1.1(cre)Thor/J) were obtained from the Jackson Laboratories. TMEM219flox/flox were generated in collaboration with Applied StemCell (Milpitas, CA) and housed at Charles River Laboratories (Wilmington, MA). TMEM219flox/floxINS1cre male mice (Beta-TMEM219−/−) were obtained by breeding TMEM219flox/flox mice with INS1cre mice, and the resulting colony was housed at Charles River Laboratories. Mice were housed at a temperature between 21 and 23 °C with 50–60% humidity and kept on a 12/12 h light/dark cycle with free access to food and water. All mice were cared for and used in accordance with institutional guidelines approved by the Boston Children’s Hospital Institutional Animal Care and Use Committee or in accordance with Italian law on animal care N° 116/1992 and the European Communities Council Directive EEC/609/86. All animal studies were approved by the Italian Ministry of Health and Local University of Milan Committee and by the Boston Children’s Hospital Institutional Animal Care and Use Committee. Some studies were conducted at the Jackson Laboratories.
D’Addio F., Maestroni A., Assi E., Ben Nasr M., Amabile G., Usuelli V., Loretelli C., Bertuzzi F., Antonioli B., Cardarelli F., El Essawy B., Solini A., Gerling I.C., Bianchi C., Becchi G., Mazzucchelli S., Corradi D., Fadini G.P., Foschi D., Markmann J.F., Orsi E., Škrha J J.r., Camboni M.G., Abdi R., James Shapiro A.M., Folli F., Ludvigsson J., Del Prato S., Zuccotti G, & Fiorina P. (2022). The IGFBP3/TMEM219 pathway regulates beta cell homeostasis. Nature Communications, 13, 684.
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2

Modulation of Slc12a8 Expression in NIH3T3 Cells and Intestinal Tissues

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NIH3T3 cells (originally purchased from the American Type Culture Collection [ATCC]) were cultured at 37 °C and 5% CO2 in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 10% fetal bovine serum (FBS), 100 units/ml penicillin, and 100 μg/ml streptomycin. For Slc12a8 mRNA expression analysis, 2.5 × 105 cells per well were incubated in 6-well plates with DMEM with 1% FBS containing 0.1% DMSO or 100 nM FK866 or 100 nM FK866 plus 100 μM NMN for 24h. Small intestines from 3 month-old C57BL/6J (B6) male mice (Jackson Laboratories) were cut into three segments with duodenum/jejunum/ileum length ratios of 1:3:2 36 (link). One centimeter of each segment was opened longitudinally, washed once with cold PBS, and incubated for 4h at 37°C with 0.1% DMSO or 100 nM FK866 or 100 nM FK866 plus 500 μM NMN in the 1:1 mixture of DMEM and Ham’s F-12 medium (Sigma) with 5% FBS and the following additives: 5 μg/ml insulin (Sigma), 20 ng/ml epidermal growth factor (Sigma), 1x B27 supplement (GIBCO), 1 mM Sodium pyruvate (Corning), 100 units/ml penicillin, 100 μg/ml streptomycin, and 2 mM glutamax (GIBCO). Cellular and tissue total RNA samples, which were extracted using the PureLink RNA Mini kit (Cat# 12183025, Ambion), were analyzed by quantitative RT-PCR, and relative expression levels were calculated for each gene by normalizing to Gapdh expression levels 16 (link).
Grozio A., Mills K.F., Yoshino J., Bruzzone S., Sociali G., Tokizane K., Lei H.C., Cunningham R., Sasaki Y., Migaud M.E, & Imai S.I. (2019). Slc12a8 is a nicotinamide mononucleotide transporter. Nature metabolism, 1(1), 47-57.
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3

Transcriptomic Profiling of FK866 Effects

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Total RNA was isolated from primary hepatocytes, pancreatic islets, and hippocampal neurospheres, treated with 0.1% DMSO (control) or FK866 (200 nM for primary hepatocytes, and 10 nM for pancreatic islets and hippocampal neurospheres). Primary hepatocytes and pancreatic islets were isolated from 3 month-old C57BL/6J (B6) male mice (Jackson Laboratories). To determine transcriptional changes induced by FK866 treatment, microarray analyses were conducted using the Illumina Mouse Ref 8 whole genome microarrays (version 2). The background-subtracted raw microarray data were subjected to Z score transformation, and Z ratios were calculated as described previously 5 (link). All data were analyzed by the R statistical software package.
Grozio A., Mills K.F., Yoshino J., Bruzzone S., Sociali G., Tokizane K., Lei H.C., Cunningham R., Sasaki Y., Migaud M.E, & Imai S.I. (2019). Slc12a8 is a nicotinamide mononucleotide transporter. Nature metabolism, 1(1), 47-57.
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4

Modulation of Slc12a8 Expression in NIH3T3 Cells and Intestinal Tissues

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NIH3T3 cells (originally purchased from the American Type Culture Collection [ATCC]) were cultured at 37 °C and 5% CO2 in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 10% fetal bovine serum (FBS), 100 units/ml penicillin, and 100 μg/ml streptomycin. For Slc12a8 mRNA expression analysis, 2.5 × 105 cells per well were incubated in 6-well plates with DMEM with 1% FBS containing 0.1% DMSO or 100 nM FK866 or 100 nM FK866 plus 100 μM NMN for 24h. Small intestines from 3 month-old C57BL/6J (B6) male mice (Jackson Laboratories) were cut into three segments with duodenum/jejunum/ileum length ratios of 1:3:2 36 (link). One centimeter of each segment was opened longitudinally, washed once with cold PBS, and incubated for 4h at 37°C with 0.1% DMSO or 100 nM FK866 or 100 nM FK866 plus 500 μM NMN in the 1:1 mixture of DMEM and Ham’s F-12 medium (Sigma) with 5% FBS and the following additives: 5 μg/ml insulin (Sigma), 20 ng/ml epidermal growth factor (Sigma), 1x B27 supplement (GIBCO), 1 mM Sodium pyruvate (Corning), 100 units/ml penicillin, 100 μg/ml streptomycin, and 2 mM glutamax (GIBCO). Cellular and tissue total RNA samples, which were extracted using the PureLink RNA Mini kit (Cat# 12183025, Ambion), were analyzed by quantitative RT-PCR, and relative expression levels were calculated for each gene by normalizing to Gapdh expression levels 16 (link).
Grozio A., Mills K.F., Yoshino J., Bruzzone S., Sociali G., Tokizane K., Lei H.C., Cunningham R., Sasaki Y., Migaud M.E, & Imai S.I. (2019). Slc12a8 is a nicotinamide mononucleotide transporter. Nature metabolism, 1(1), 47-57.
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5

Transcriptomic Profiling of FK866 Effects

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Total RNA was isolated from primary hepatocytes, pancreatic islets, and hippocampal neurospheres, treated with 0.1% DMSO (control) or FK866 (200 nM for primary hepatocytes, and 10 nM for pancreatic islets and hippocampal neurospheres). Primary hepatocytes and pancreatic islets were isolated from 3 month-old C57BL/6J (B6) male mice (Jackson Laboratories). To determine transcriptional changes induced by FK866 treatment, microarray analyses were conducted using the Illumina Mouse Ref 8 whole genome microarrays (version 2). The background-subtracted raw microarray data were subjected to Z score transformation, and Z ratios were calculated as described previously 5 (link). All data were analyzed by the R statistical software package.
Grozio A., Mills K.F., Yoshino J., Bruzzone S., Sociali G., Tokizane K., Lei H.C., Cunningham R., Sasaki Y., Migaud M.E, & Imai S.I. (2019). Slc12a8 is a nicotinamide mononucleotide transporter. Nature metabolism, 1(1), 47-57.
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6

Alcohol Drinking in C57BL/6J Mice

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This study used 60 adult C57BL/6J (B6) male mice that were 8 weeks of age at onset of drinking and weighed 25-30g (Jackson Laboratories, Sacramento, CA). Animals were randomly divided into an alcohol-drinking group (n=30; hereafter referred to as Alcohol mice) and a water-drinking group (n=30; hereafter referred to as Water mice) and then individually housed in standard, Plexiglas cages, under a 12-hour-reverse light/dark cycle (lights off at 10am), in a temperature-controlled vivarium (23°C). Food and water were available ad libitum, with the exception of the 2-hr alcohol drinking period, during which time the home cage water bottle was removed. All experiments were conducted in compliance with the National Institutes of Health Guide for Care and Use of Laboratory Animals (NIH Publication No. 80–23, revised 2010) and approved by the IACUC of the University of California, Santa Barbara.
Lee K.M., Coehlo M., McGregor H.A., Waltermire R.S, & Szumlinski K.K. (2015). Binge Alcohol Drinking Elicits Persistent Negative Affect in Mice. Behavioural brain research, 291, 385-398.
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7

Voluntary Ethanol Consumption in B6 Mice

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All studies were conducted in adult drug-naïve C57BL/6J (B6) male mice that were purchased at 8 weeks of age from Jackson Laboratories (Bar Harbor, ME). We chose this strain because of their propensity for voluntary ethanol consumption 6 (link). Seventy-two mice between 8 and 12 weeks old were used: 32 for the ethanol experiment in which mice received ethanol and PPAR agonists or saline (4 groups with 8 mice each) and 40 for the microarray experiment in which mice received PPAR agonists or saline only (4 groups with 10 mice each). Mice were housed in the Animal Resources Center at The University of Texas at Austin with 12-h light/dark cycles (lights on at 10:00 AM) and had ad libitum access to rodent chow (Prolab RMH 180 5LL2 chow, TestDiet, Richmond, IN) and water. All efforts were made to minimize animal suffering, to reduce the number of animals used, and to utilize alternatives to in vivo techniques, if available. All experiments were approved by The University of Texas at Austin Institute for Animal Care and Use Committee and were conducted in accordance with NIH guidelines with regard to the use of animals in research.
Ferguson L.B., Most D., Blednov Y.A, & Harris R.A. (2014). PPAR agonists regulate brain gene expression: Relationship to their effects on ethanol consumption. Neuropharmacology, 86, 397-407.
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8

Aging and Gut Microbiome in Mice

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Adult C57BL/6J (B6) male mice were purchased at ages 6–8 weeks and 18–20 months from Jackson Laboratory (Bar Harbor, ME). Prior to initiation of the experiment, mice were acclimated to the humidity-controlled housing room programmed for a 12-hour light-dark cycle for a minimum of 2 weeks. Mice were cared for by the University of Florida Animal Care Services (Gainesville, FL). Mice of the same age cohort and treatment were housed in transparent cages (3–4 animals per cage) within specific pathogen-free facilities at ambient room temperatures (23°C). The animals were provided standard rodent chow and water ad libitum for the duration of the study. Due to the coprophagic nature of mice, caging them together for 2 weeks allowed the mice to obtain a similar microbiota composition and structure (18 (link), 19 (link)), although our previous worked revealed that microbiome similarity occurred only among mice of the same age (20 (link)). The animals were cared for and used according to the Guide for the Care and Use of Laboratory Animals, and the experiments were approved by the University of Florida Institutional Animal Care and Use Committee.
Darden D.B., Mira J.C., Lopez M.C., Stortz J.A., Fenner B.P., Kelly L.S., Nacionales D.C., Budharaju A., Loftus T.J., Baker H.V., Moore F.A., Brakenridge S.C., Moldawer L.L., Mohr A.M, & Efron P.A. (2021). Identification of Unique microRNA Expression Patterns in Bone Marrow Hematopoietic Stem and Progenitor Cells after Hemorrhagic Shock and Polytrauma in Young and Old Adult Mice. The journal of trauma and acute care surgery, 91(4), 692-699.
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