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Then cells were washed in PBS and stimulated with RPMI 1640 supplemented with 100 U/ml penicillin, and 100 µg/ml streptomycin, 10% (v/v) heat-inactivated fetal bovine serum, 2 mM L-Glutamine, 2, 5 or 16 mM glucose for 24 hours in replicas of 6 followed by RNA extraction.
For INS-1 cells treatments were given for 1 and 24 hours, in replicas of 6.
For INS-1E and INS-1E-ZnT8-EGFP cells treatments were given for 6 and 24 hours in replicas of 3. Controls were incubated with RPMI 1640 medium and 11 mM glucose.
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Then cells were stimulated for 24 hours with 2, 5 or 16 mM glucose in replicas of 9 followed by RNA extraction.
Controls were stimulated for 24 hours with RPMI 1640 medium and 5 mM glucose in replicas of 6 and 3 for RNA extraction and insulin measurements, respectively.
While Mr. Bush's advisers said the president spent much of this week in informal, conversational sessions simply tossing lines back and forth, Mr. Kerry practiced for hours with replicas of the lights, developing four or five ways to sum up quickly.
Fortunately, some bento bloggers are already fighting the impulse to spend hours making perfect replicas of Hello Kitty.
In particular, we focused on DM-CGIs found in culture replicas of the same cell line.
In replica exchange, replicas of the system are simulated in parallel at different temperatures.
I spent a couple of hours in his apartment, then I made a replica and practised with the replica at home for a week.
Five years and a million volunteer hours in the building, this handsome replica was launched in 2000 as an educational vessel and flagship for the state of Wisconsin.
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