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Cytochrome P450 isoforms 1A2, 2C9, 2C19, 2D6, and 3A4 were obtained as singly expressed enzymes (Baculovirus transfected insect cell system, rhCYPs) from BD Gentest (Woburn, MA, USA).
Cytosolic fractions containing singly expressed CFP, and YFP, respectively, were mixed in different ratios.
This phenomenon was evaluated in cells expressing only one of the two constructs, because the contribution of CFP and YFP to FRET in coexpressed cells is proportional to that in singly expressed cells.
First, we assessed the subcellular localization of RPGR1 19 or RPGRORF15 when singly expressed.
Therefore, APRIL seems not to be co-precipitated with any singly expressed vRdRP subunit (Watanabe et al., 2014).
Moreover, these mutations affect the subcellular partitioning of RPGRIP1α1 when singly expressed at reduced levels in cells (Lu et al., 2005).
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In fact, it was observed that plants singly expressing bgl1C, cel6B, cel9A, and xeg74 genes from T. fusca showed severe pleiotropic effects [ 25].
Moreover, in cells expressing both PrPC and Dpl, cholesterol depletion was not able to induce the dissociation of the two proteins from DRMs (from 62±2.4 to 60±1.5% for GFP Dpl and from 36±1.8 to 33±1.5% for PrPC), which was observed in singly expressing clones (from 65±5.3 to 35±2.4% of GFP Dpl and from 60±2.1 to 30±1.3% of PrPC; Figure 4).
As shown in Fig 4C, Trim32 mainly resided in cytoplasmic small granules in Trim32-singly expressed cells.
To confirm the predicted interaction between LZAP and p38, Myc-tagged LZAP was transiently expressed singly or with Flag-tagged p38 in mammalian U2OS cells before immunoprecipitation.
When the proteins are expressed singly, GBP-3 has been shown to localize in the cytoplasm in a diffuse manner whereas GBP-4 was evenly distributed throughout the nucleus and cytoplasm [11].
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