Words similar to egfp-cpla

Example sentences for: egfp-cpla

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2 , it is necessary to over-express the enzyme several fold as previously reported [ 16 ] . Cytosolic PLA 2 has been shown to mediate Ca 2+-induced AA release in MDCK cells treated with ATP and IONO in experiments using the group IV cPLA 2 α-specific inhibitor pyrrolidine-1 [ 27 ] . To measure cPLA 2 mediated AA release, EGFP-cPLA 2 -transfected MDCK cells labeled with [ 3H]-AA were incubated with 0.3, 1 or 10 μM U0126 for 15 min prior to stimulation with 100 μM ATP, 1 μM IONO, or 10 μM IONO.
Stable lines expressing EGFP-cPLA 2 were generated by growing transfected cells in growth medium for 3 d, supplementing the growth medium with 5 mg/ml Geneticin (antibiotic G418-sulfate), and culturing for an additional 2 wk in Geneticin.
However, EGFP-cPLA 2 is functional since it dose-dependently catalyzes release of AA when expressed in cells that lack endogenous cPLA 2 , such as Sf9 cells [ 13 ] and immortalized mouse lung fibroblasts from cPLA 2 α knock-out mice [ 26 ] . In order to enhance AA release in cells containing endogenous cPLA
Stable EGFP-cPLA 2 transfectants were grown on 100 mm dishes at 5 × 10 3cells/cm 2in growth medium for one day, then quiesced in serum-free medium overnight.
EGFP-cPLA 2 was expressed to similar levels as endogenous enzyme but did not contribute significantly to AA release in stably transfected cells.