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blot analysis\nProtein extracts were isolated from homogenized liver (freshly harvested and flash frozen) of wildtype, Line 525A, and Line A transgenic mice. Protein (25 μg per sample) was analyzed by electrophoresis on a 10% SDS-PAGE Ready Gel (Bio-Rad, Hercules, CA) and transferred to PVDF membrane (Sequi-Blot; Bio-Rad) via electroblotting. All blots were probed for E2 protein using polyclonal rabbit E2 antisera (1:5,000), which detects both mouse (~47 kD) and human (~54 kD) E2 subunits ([30]; gift from Dr. Susan Hutson, Wake Forest University). Blots were re-probed with a rabbit anti-c-myc tag antibody (1:10,000; abCam, Cambridge, MA; cat.# ab9106-100). Blots were also re-probed with an antibody for β-actin (43 kD; 1:10,000; abCam; cat.# ab8227-50) to allow for loading comparisons. A goat anti-rabbit secondary antibody conjugated to horseradish peroxidase (1:10,000; Novus, Littleton, CO; cat.# NB730-H) was used for detection using the Western Lightning chemiluminescence reagent (Perkin Elmer, Boston, MA) and exposed to X-ray film."}

{"project":"craft-ca-core-ex-dev","denotations":[{"id":"T8787","span":{"begin":22,"end":29},"obj":"CHEBI_PR_EXT:protein"},{"id":"T8788","span":{"begin":70,"end":75},"obj":"UBERON:0002107"},{"id":"T8789","span":{"begin":116,"end":124},"obj":"SO_EXT:wild_type_entity_or_quality"},{"id":"T8790","span":{"begin":148,"end":158},"obj":"SO_EXT:transgenic_entity"},{"id":"T8791","span":{"begin":159,"end":163},"obj":"NCBITaxon:10088"},{"id":"T8792","span":{"begin":165,"end":172},"obj":"CHEBI_PR_EXT:protein"},{"id":"T8793","span":{"begin":233,"end":236},"obj":"CHEBI:8984"},{"id":"T8794","span":{"begin":280,"end":291},"obj":"GO_EXT:biological_movement_or_translocation_process"},{"id":"T8795","span":{"begin":295,"end":299},"obj":"CHEBI:53250"},{"id":"T8796","span":{"begin":378,"end":380},"obj":"PR_EXT:000006300"},{"id":"T8797","span":{"begin":381,"end":388},"obj":"CHEBI_PR_EXT:protein"},{"id":"T8798","span":{"begin":406,"end":412},"obj":"NCBITaxon:9986"},{"id":"T8799","span":{"begin":413,"end":415},"obj":"PR_EXT:000006300"},{"id":"T8800","span":{"begin":420,"end":424},"obj":"UBERON:0001977"},{"id":"T8801","span":{"begin":455,"end":460},"obj":"NCBITaxon:10088"},{"id":"T8802","span":{"begin":474,"end":479},"obj":"NCBITaxon:9606"},{"id":"T8803","span":{"begin":489,"end":500},"obj":"PR_EXT:000006300"},{"id":"T8804","span":{"begin":589,"end":595},"obj":"NCBITaxon:9986"},{"id":"T8805","span":{"begin":601,"end":606},"obj":"PR_EXT:000000084"},{"id":"T8806","span":{"begin":607,"end":610},"obj":"CHEBI_SO_EXT:molecular_indicator_or_label_or_marker_or_tag"},{"id":"T8807","span":{"begin":611,"end":619},"obj":"GO:0042571"},{"id":"T8808","span":{"begin":706,"end":714},"obj":"GO:0042571"},{"id":"T8809","span":{"begin":719,"end":726},"obj":"PR_EXT:000003676"},{"id":"T8810","span":{"begin":805,"end":809},"obj":"NCBITaxon:9925"},{"id":"T8811","span":{"begin":815,"end":821},"obj":"NCBITaxon:9986"},{"id":"T8812","span":{"begin":832,"end":840},"obj":"GO:0042571"},{"id":"T8813","span":{"begin":841,"end":851},"obj":"MOP:0000779"},{"id":"T8814","span":{"begin":855,"end":866},"obj":"NCBITaxon:3704"},{"id":"T8815","span":{"begin":867,"end":877},"obj":"GO_EXT:0004601"},{"id":"T8816","span":{"begin":995,"end":1002},"obj":"CHEBI_EXT:33893"}],"text":"Western blot analysis\nProtein extracts were isolated from homogenized liver (freshly harvested and flash frozen) of wildtype, Line 525A, and Line A transgenic mice. Protein (25 μg per sample) was analyzed by electrophoresis on a 10% SDS-PAGE Ready Gel (Bio-Rad, Hercules, CA) and transferred to PVDF membrane (Sequi-Blot; Bio-Rad) via electroblotting. All blots were probed for E2 protein using polyclonal rabbit E2 antisera (1:5,000), which detects both mouse (~47 kD) and human (~54 kD) E2 subunits ([30]; gift from Dr. Susan Hutson, Wake Forest University). Blots were re-probed with a rabbit anti-c-myc tag antibody (1:10,000; abCam, Cambridge, MA; cat.# ab9106-100). Blots were also re-probed with an antibody for β-actin (43 kD; 1:10,000; abCam; cat.# ab8227-50) to allow for loading comparisons. A goat anti-rabbit secondary antibody conjugated to horseradish peroxidase (1:10,000; Novus, Littleton, CO; cat.# NB730-H) was used for detection using the Western Lightning chemiluminescence reagent (Perkin Elmer, Boston, MA) and exposed to X-ray film."}

{"project":"craft-ca-core-dev","denotations":[{"id":"T8765","span":{"begin":70,"end":75},"obj":"UBERON:0002107"},{"id":"T8766","span":{"begin":159,"end":163},"obj":"NCBITaxon:10088"},{"id":"T8767","span":{"begin":233,"end":236},"obj":"CHEBI:8984"},{"id":"T8768","span":{"begin":295,"end":299},"obj":"CHEBI:53250"},{"id":"T8769","span":{"begin":378,"end":380},"obj":"PR:000006300"},{"id":"T8770","span":{"begin":406,"end":412},"obj":"NCBITaxon:9986"},{"id":"T8771","span":{"begin":413,"end":415},"obj":"PR:000006300"},{"id":"T8772","span":{"begin":420,"end":424},"obj":"UBERON:0001977"},{"id":"T8773","span":{"begin":455,"end":460},"obj":"NCBITaxon:10088"},{"id":"T8774","span":{"begin":474,"end":479},"obj":"NCBITaxon:9606"},{"id":"T8775","span":{"begin":489,"end":500},"obj":"PR:000006300"},{"id":"T8776","span":{"begin":589,"end":595},"obj":"NCBITaxon:9986"},{"id":"T8777","span":{"begin":601,"end":606},"obj":"PR:000000084"},{"id":"T8778","span":{"begin":611,"end":619},"obj":"GO:0042571"},{"id":"T8779","span":{"begin":706,"end":714},"obj":"GO:0042571"},{"id":"T8780","span":{"begin":719,"end":726},"obj":"PR:000003676"},{"id":"T8781","span":{"begin":805,"end":809},"obj":"NCBITaxon:9925"},{"id":"T8782","span":{"begin":815,"end":821},"obj":"NCBITaxon:9986"},{"id":"T8783","span":{"begin":832,"end":840},"obj":"GO:0042571"},{"id":"T8784","span":{"begin":841,"end":851},"obj":"MOP:0000779"},{"id":"T8785","span":{"begin":855,"end":866},"obj":"NCBITaxon:3704"},{"id":"T8786","span":{"begin":995,"end":1002},"obj":"CHEBI:33893"}],"text":"Western blot analysis\nProtein extracts were isolated from homogenized liver (freshly harvested and flash frozen) of wildtype, Line 525A, and Line A transgenic mice. Protein (25 μg per sample) was analyzed by electrophoresis on a 10% SDS-PAGE Ready Gel (Bio-Rad, Hercules, CA) and transferred to PVDF membrane (Sequi-Blot; Bio-Rad) via electroblotting. All blots were probed for E2 protein using polyclonal rabbit E2 antisera (1:5,000), which detects both mouse (~47 kD) and human (~54 kD) E2 subunits ([30]; gift from Dr. Susan Hutson, Wake Forest University). Blots were re-probed with a rabbit anti-c-myc tag antibody (1:10,000; abCam, Cambridge, MA; cat.# ab9106-100). Blots were also re-probed with an antibody for β-actin (43 kD; 1:10,000; abCam; cat.# ab8227-50) to allow for loading comparisons. A goat anti-rabbit secondary antibody conjugated to horseradish peroxidase (1:10,000; Novus, Littleton, CO; cat.# NB730-H) was used for detection using the Western Lightning chemiluminescence reagent (Perkin Elmer, Boston, MA) and exposed to X-ray film."}