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Insulin glargine was given to female rats before mating, during mating, and throughout pregnancy at doses up to 0.36 mg/kg/day, which is approximately 50 times the recommended human subcutaneous starting dosage of 0.2 Units/kg/day (0.007 mg/kg/day)

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After discarding the supernatant, the combined pellet was resuspended in 25 ml of ice-cold PBS buffer

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