Mouse strains were purchased from The Jackson Laboratory or received as gifts and are referenced in Extended Data Table 1 . Mice, maintained under specific pathogen–free conditions, were handled in accordance with protocols approved and monitored by the Dana-Farber Cancer Institute Animal Care and Use Committee. Male and female C57BL/6 mice were used between 2 and 6 months of age. Dibenzazepine (DBZ, Syncom, Groningen, The Netherlands) was suspended in 0.5% (w/v) hydroxypropylmethyl-cellulose (Methocel E4M, Dow Chemicals) and 0.1% (w/v) Tween 80 (Bio-Rad) in water and 100 μmol/kg was administered by intraperitoneal (IP) injection. We activated Cre-ER recombinase by IP injection of 1 mg tamoxifen (Invitrogen) for 5 days or one dose of 4 mg tamoxifen. Cre-PR recombinase was induced by 2 IP injections of 2 mg RU486 (Sigma) approximately 8 h apart. BrdU (B-D Biosciences, 50 mg/kg) was injected IP and mice were sacrificed 1 h or 3 days later.
Methocel e4m
Manufactured by Dow
Methocel E4M is a cellulose-based product manufactured by Dow. It functions as a thickening, suspending, and film-forming agent for various applications.
Automatically generated - may contain errors
8 protocols using methocel e4m
1
Modulating Mouse Genetic Pathways
2
Adipose Tissue Deposition of PLGA Microspheres
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Mice were first anesthetized by a ketamine–xylazine cocktail and then either FITC-dextran-loaded PLGA microspheres or DBZ-loaded PLGA microspheres (20 mg/30 g body weight) were injected into the inguinal WAT depot in one side of the body in a 1 mL solution of 0.5% Methocel E4M (wt/vol; Dow Chemical) and 0.1% Tween-80 (wt/vol; Sigma) in water. Similarly, PLGA microspheres were injected into the contralateral inguinal WAT depot. Adipose tissues were harvested after 24 h and 14 days post-injection for the study with FITC-dextran-loaded PLGA microspheres and DBZ-loaded PLGA microspheres, respectively. Oil Red O staining was performed to label the adipocytes for the study with FITC-dextran-loaded PLGA microspheres whereas hematoxylin and eosin (H&E) and immunohistochemistry staining was conducted for the study with DBZ-loaded PLGA microspheres.
3
CDK9 Inhibitors Modulate Arthritis in Mice
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DBA/1 mice were treated with specific CDK9 inhibitors, PC585 and PC579 at 10 mg/kg. The inhibitors were sonicated in 0.8% methyl cellulose (Methocel E4M, Dow) and provided to mice through oral gavage. i.p injections of 4 mg/kg Enbrel (etanercept, Wyeth Europe) dosed biweekly was used as a reference. Treatment started three days prior induction of arthritis. For the mRNA expression study daily treatment with 10 mg/kg of PC585 was initiated at onset of arthritis and sustained for 21 days, until end of experiment. Healthy NMRI mice were treated for 7 consecutive days with PC585 to assess the effect of treatment on T cell populations.
4
Xenograft Model of Tumor Growth
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2 × 106 human LPS246 cells were injected subcutaneously into the flank of 16–17-wk-old NSG mice. Before injection, the cells were suspended in 0.1 ml HBSS and mixed with 0.1 ml Matrigel. DBZ was purchased from TOCRIS Bioscience and dissolved in DMSO at 100 mM concentration. Immediately before administration, the stock was suspended at 1:100 dilution in a solution containing 0.5% Methocel E4M (wt/vol; Dow Chemical) and 0.1% Tween-80 (wt/vol; Sigma-Aldrich) in H2O. The working solution was mixed by vortex for 1 min, and i.p. injected at a dosage of 10 µmol DBZ/kg mouse body weight every 2 d starting from day 14 after LPS246 cell injection. Control group mice were injected with equal volumes of DMSO diluted in E4M/Tween-80 solution. Control and DBZ treatment groups were randomly grouped.
5
Xenograft Tumor Growth Inhibition
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Human LPS246 cells (2 × 106) were injected subcutaneously into the flank of 16–17-wk-old NSG mouse. Before injection, the cells were suspended in 50 µl HBSS and mixed with 50 µl Matrigel. DBZ was purchased from TOCRIS Bioscience and dissolved in DMSO at a 100-mM concentration. Immediately before administration, the stock was suspended at 1:100 dilution in a solution containing 0.5% Methocel E4M (wt/vol; Dow Chemical) and 0.1% Tween-80 (wt/vol; Sigma-Aldrich) in H2O. The working solution was mixed by vortex and sonication for 1 min, and i.p. injected at a dosage of 10 µmol DBZ/kg mouse body weight every 2 d starting from day 15 after LPS246 cell injection. Control group mice were injected with equal volumes of DMSO diluted in E4M/Tween-80 solution. Control and DBZ treatment groups were randomly grouped.
6
Modulating Mouse Genetic Pathways
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Mouse strains were purchased from The Jackson Laboratory or received as gifts and are referenced in Extended Data Table 1 . Mice, maintained under specific pathogen–free conditions, were handled in accordance with protocols approved and monitored by the Dana-Farber Cancer Institute Animal Care and Use Committee. Male and female C57BL/6 mice were used between 2 and 6 months of age. Dibenzazepine (DBZ, Syncom, Groningen, The Netherlands) was suspended in 0.5% (w/v) hydroxypropylmethyl-cellulose (Methocel E4M, Dow Chemicals) and 0.1% (w/v) Tween 80 (Bio-Rad) in water and 100 μmol/kg was administered by intraperitoneal (IP) injection. We activated Cre-ER recombinase by IP injection of 1 mg tamoxifen (Invitrogen) for 5 days or one dose of 4 mg tamoxifen. Cre-PR recombinase was induced by 2 IP injections of 2 mg RU486 (Sigma) approximately 8 h apart. BrdU (B-D Biosciences, 50 mg/kg) was injected IP and mice were sacrificed 1 h or 3 days later.
7
DBZ Treatment for Genetic Obesity
8
DBZ Treatment for Genetic Obesity
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DBZ was purchased from TOCRIS Bioscience (Cat. No. 4489) and used following previous studies45 (link),63 (link). DBZ was dissolved in DMSO at 100 mM concentration. Upon use the stock was suspended at 1:100 dilution in a solution containing 0.5% Methocel E4M (w/v, Dow Chemical) and 0.1% Tween-80 (w/v Sigma) in H2O. This working solution was mixed by vortex and sonication for 1 minute each, and IP injected at a dosage of 10 μmol DBZ per kg body weight. Control groups were injected with equal volumes of DMSO diluted in E4M/Tween-80 solution. Control and DBZ treatment groups were randomly grouped. To test the effect of DBZ on genetic induced obesity, ob/ob (Lepob) mice (6-week-old, male) were treated every other day for one month.
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