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When a culture attained steady state (after 5 volume changes), samples were collected for estimation of dry cell mass, fluoride release and residual 4-FCA.
For platelet identification and glycoprotein IIb/IIIa (GPIIb/IIIa) expression changes samples were stained with 6 μl anti-GPIIb/IIIa PE (phycoerythin -conjugated monoclonal antibody (CD41-phycoerythin -conjugatedulter, Krefeld, Germonoclonal
For experiment 3 (RT-qPCR), both symbiotic and aposymbiotic anemones were maintained at 25° on a 12L:12D cycle with feeding every 2 d, followed by water changes; samples were collected 2 d after the last feeding and 6 hr into the light period.
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To observe the temporal change, samples were collected after heating for 30, 60, and 300 s.
The structure of the phase-change samples was checked by X-ray diffraction [8].
Thermal stabilities and weight changes of samples were tested by TGA.
To identify structural changes, some samples were tempered at 600 or 640 °C.
Histological changes in samples were examined by light microscopy.
Fold changes between samples were determined using the ΔΔCt method.
Gene and transcript expression changes among samples were analyzed with Cuffdiff software.
To identify hormone refractory dependent expression changes MET_HR samples were scaled to the median across the non-refractory samples MET_HN.
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