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To control for transfection efficiency, the vector pLP-ECFP C1 that expresses ECFP was co-transfected with the test expression vectors, and the amount of fluorescence in the cell lysates was measured.
After the monkeys established morphine CPP, their insulae were reversibly inactivated by bilateral microinjection with 5% lidocaine (40 μl) prior to the post-conditioning test (expression) of CPP using a within-subject design.
One ml of culture was removed every 24 h to test expression levels using the synthetic 4-nitrophenyl β-xylopyranoside (PNPX) as described in section 2.4.
RNA was isolated from brain, liver and skeletal muscle with Trizol reagent (Invitrogen) to test expression.
Therefore, we sought to test expression of IK2 in puf2 sporozoites, in comparison to WT and puf1 sporozoites, using RT-qPCR.
Given that Scarb1 was an obvious candidate, real-time PCR was performed using liver tissue to test expression of Scarb1 between HLB398 mutant mice and B6 controls.
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The test expressions of the mutants were analyzed following lysis and centrifugation.
Crude extracts from test expressions were centrifuged at 4°C at 16,000 g for 60 minutes.
Test expressions at 25°C had no observable effect and resulted in insoluble protein (not shown).
Subsequently, E. coli BL21 DE3) as well as E. coli BL21 DE3 pLysS were used as host strains for test expressions.
We tested expression of Turbo, tdTomato, mCherry, mKat and mPlum expressed from Phsp60 or Psmyc.
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