The TLR4 floxed mice (B6(Cg)-Tlr4tm1.1Karp/J mice) and B6 FVB-Tg (cdh5-cre)7Mlia/J Cre mice were purchased from Jackson Laboratories. After 2 generations, TLR4 floxed mice were bred with cdh5-Cre mice to generate conditional knockout mice in which TLR4 is eliminated in vascular endothelial cells. Around 3 months of age, TLR4 floxed and TLR4 Cre-Lox mice were used for experiments.
B6 fvb tg cdh5 cre 7mlia j cre mice
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B6 FVB-Tg (cdh5-cre)7Mlia/J Cre mice express Cre recombinase under the control of the endothelial-specific cadherin 5 (Cdh5) promoter. This allows for Cre-mediated recombination in endothelial cells.
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13 protocols using b6 fvb tg cdh5 cre 7mlia j cre mice
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Endothelial-specific TLR4 Knockout Mice
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Conditional Endothelial Epac1 Knockout Mice
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Epac1 floxed mice (B6;129S2-Rapgef3tm1Geno/J mice) and B6 FVB-Tg (cdh5-cre)7Mlia/J Cre mice were purchased from Jackson Laboratories. After 2 generations, Epac1 floxed mice were bred with cdh5-Cre mice to generate conditional knockout mice in which Epac1 is eliminated in endothelial cells. We have previously reported studies using these mice. In our previous studies, we showed successful loss of Epac1 in the Epac1 EC-KO mice using genotyping, Western blotting of whole retinal lysates, and immunostaining of 10um retinal sections. The retinal sections were stained with isolectin B4 (endothelial cells) and Epac1 (red) to show little co-localization of Isolectin GS-IB4 and Epac1 in the Epac1 EC-KO mice in blood vessels (Fig 2 of previous work [12 (link)]). Mice of both sexes at 2 months of age were used for these studies. All work was reviewed and approved by the Institutional Animal Care and Use Committee of Wayne State University School of Medicine (Protocol #17-07-301) and conforms to NIH guidelines.
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Conditional Knockout of TLR4 in Vascular Endothelial Cells
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Conditional Epac1 Knockout in Vascular Endothelial Cells
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Animal procedures meet the Association for Research in Vision and Ophthalmology requirements and were approved by the Institutional Animal Care and Use Committee of Wayne State University and conform to NIH guidelines. Epac1 floxed mice (B6;129S2-Rapgef3tm1Geno/J mice) and B6 FVB-Tg (cdh5-cre)7Mlia/J Cre mice were purchased from Jackson Laboratories. The Epac1 floxed mice were bred with the cdh5-Cre mice to generate conditional knockout mice in which Epac1 is eliminated in vascular endothelial cells. At 3 months of age, Epac1 floxed and Epac1 Cre-Lox mice were used for these experiments.20 (link),21 (link) Euthanasia was performed with drug overdose followed by cervical dislocation. Whole retinal lysates were collected from the mice.
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Epac1 Knockout in Vascular Endothelial Cells
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All animal procedures met the Association for Research in Vision and Ophthalmology requirements and were approved by the Institutional Animal Care and Use Committee of Wayne State University and conformed to NIH guidelines. Epac1 floxed mice (B6; 129S2-Rapgef3tm1Geno/J mice) and B6 FVB-Tg (cdh5-cre)7Mlia/J Cre mice were purchased from Jackson Laboratories. After 2 generations, the Epac1 floxed mice were bred with cdh5-Cre mice to generate conditional knockout mice in which Epac1 is eliminated in vascular endothelial cells [15 (link)]. At 3 months of age, Epac1 floxed and Epac1 Cre-Lox mice were used for experiments.
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Conditional Knockdown of Epac1 in Vascular Endothelium
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Epac1 floxed mice (B6;129S2-Rapgef3tm1Geno/J mice) and B6 FVB-Tg (cdh5-cre)7Mlia/J Cre mice were purchased from Jackson Laboratories (Bar Harbor, ME). Epac1 floxed mice were bred with cdh5-Cre mice to generate a line of conditional knockout mice with Epac1 eradicated from vascular endothelial cells [12 (link)]. Retinal lysates from Epac1 floxed and Epac1 Cre-Lox mice were collected at 2 months of age. All animal procedures met the Association for Research in Vision and Ophthalmology requirements, were approved by the Institutional Animal Care and Use Committee of Wayne State University, and conformed to NIH guidelines.
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Conditional miR-15a/16 Knockout in Endothelial Cells
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miR-15a/16 floxed mice (B6-129S-mirc30.tm1.1rdf/J) and B6.FVB-Tg (cdh5-cre)7Mlia/J Cre mice were purchased from Jackson Laboratories. The miR-15a/16 floxed mice were crossed with cdh5-Cre mice to generate conditional knockout mice in which miR-15a/16 is eliminated in vascular endothelial cells. At 3 months of age, both male and female mice from miR-15a/16 floxed and miR-15a/16 Cre-LoxP groups were used for further analyses. All animal procedures were reviewed and approved by the Institute Animal Care and Use Committees of the Wayne State University School of Medicine (Protocol # A 11-08-14) and conform to NIH guidelines.
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Conditional Epac1 Knockout in Endothelial Cells
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Epac1 floxed mice (B6;129S2-Rapgef3tm1Geno/J mice) and B6.FVB-Tg(Cdh5-cre)7Mlia/J Cre mice were purchased from The Jackson Laboratory (Bar Harbor, ME, USA). After two generations, Epac1 floxed mice were bred with Cdh5-cre mice, generating conditional knockout mice where Epac1 was eliminated in the vascular endothelial cells.11 (link) When they were 2 months of age, Epac1 floxed and Epac1 Cre-Lox mice were used to collect retinal samples.
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Endothelial Cell Epac1 Knockout Mice
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All animal procedures complied with the ARVO Statement for the Use of Animals in Ophthalmology and Vision Research, were approved by the Institutional Animal Care and Use Committee of Wayne State University, and conformed to National Institutes of Health guidelines (Protocol 17-07-301). Epac1 floxed mice (B6;129S2-Rapgef3tm1Geno/J mice) and B6 FVB-Tg (cdh5-cre)7Mlia/J Cre mice were purchased from The Jackson Laboratory (Bar Harbor, ME, USA). After two generations, Epac1 floxed mice were bred with cdh5-Cre mice to generate conditional knockout mice for Epac1, where Epac1 is eliminated in endothelial cells.5 Some mice were made diabetic with 60-mg/kg streptozotocin dissolved in citrate buffer, which was administered for 5 consecutive days, as we have done previously.5 Diabetes was accepted if glucose levels were greater than 250 mg/dL. Protein samples were taken from 3-month-old male and female Epac1 and cdh5/Epac1 Cre mice, and diabetic samples were collected after 2 months of diabetes in the mice. All mice were euthanized using a ketamine/xylazine overdose, followed by cervical dislocation.
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Conditional TLR4 Knockout Mouse Models
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