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At the end of the experiment, control hearts from mice that had been maintained on doxycycline were obtained.
Importantly, tTA/HIF-1α-PPN mice maintained on doxycycline showed no band on the Western blot, indicating tight control of HIF-1α-PPN expression.
Animals mated to generate RIP7-rtTA; tet-o-PyMT-IRES-Luc bitransgenic mice received dietary doxycycline to allow induction of the transgene during pancreatic organogenesis throughout embryonic development in pregnant females; the resulting bitransgenic progeny were also maintained on doxycycline.
Recipients were maintained on doxycycline to continue transgene induction.
Unless otherwise indicated, mating pairs were maintained on doxycycline (DOX -containing chow (200 mg/kg; Bio-Serve, Frenchtown, NJ) for suppression of HCV-core DOX -containingent and through weaning.
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Mice were maintained on the doxycycline supplemented diet until the end of the study.
Mice pregnant with Pdx1-tTA; tet-o-PyMT-IRES-Luc embryos and the resulting bitransgenic progeny (a cohort of 50 mice) were maintained on a doxycycline-free diet so that PyMT would be expressed throughout pancreatic development and postnatal life.
More than 90% of the Pdx1-tTA; tet-o-PyMT-IRES-Luc bitransgenic mice (45 of 50; ages between 4 months to 1 year) maintained on a doxycycline-free diet exhibited moderate luciferase activity (106 to 107 photons/sec) and had enlarged islets composed mainly of insulin-expressing β cells (Figure 2A ¸ middle row).
Cultures were maintained on puromycin and doxycycline medium, unless otherwise indicated.
Double transgenic mice (tTA/HIF-1α-PPN) were maintained on 200 µg doxycycline per ml of 2.5% sucrose-water to suppress HIF-1α-PPN expression.
To induce transgene expression, breeders and experimental mice were maintained on 2 mg/ml doxycycline (Research Products International Corp., Mount Prospect, IL, USA; 50213285).
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