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While many were localised throughout the cytoplasm in a manner similar to free eGFP, others showed more restricted localisation.
The three type D hexokinases PpHxk9, PpHxk10 and PpHxk11 also possess membrane anchors and show a similar, though more restricted localisation as the type B proteins.
One of the main characteristics of the disease is the restricted localisation of the malignant plasma cells in the BM.
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Smo-YFP distribution assessment was done in cells treated for 2 hrs with vehicle or Shh-N (6 μg/ml) prior to fixation and cells were counted and grouped into three distinct categories of localisation pattern: solely cytoplasmic (cyto), a mixed distribution of cytoplasmic and ciliary (both), and restricted ciliary localisation (primary cilium).
The only other Sub-Group with either strong support from our molecular analysis or restricted geographical localisation is Sub-Group 1.1 whose distribution is centred upon Ethiopia and Spain but whose allelic spectrum suggests that it lacks large scale introgression from any particular wild germplasm.
In an effort to try and understand if NCS-1 was unable to mediate an in vivo modulation of CN activity due to its restricted membrane localisation we also examined NFAT-GFP phosphorylation in the presence of co-expressed NCS-1G2A, a protein that fails to become myristoylated and as a result remains cytosolic.
FAPP1-PH and OSBP1-PH have also been demonstrated to bind ARF1 and thus to restrict their localisation to specific membranes through the coincident detection of both Arf1 and PtdIns4 P [8].
The GSK-3 dependent phosphorylation site S76 (Streets et al., 2006), the casine kinase 2 phosphorylation site at S812, responsible for restricting protein localisation to the ER and cilia (Cai et al., 2004), and the PKD-dependent phosphorylation site, S801 (Streets et al., 2010; all highlighted), are also not conserved in the Dictyostelium protein.
The distribution of fluorescence produced following calcein-AM addition displayed a similar pattern to doxorubicin, with localisation restricted to the outer few cell layers.
We clarified this issue by identifying and characterising a predicted C-terminal membrane helix that mediated the restricted sub-cellular localisations of CaBP7 and CaBP8.
A key feature of Ca2+-sensing proteins is that they often exhibit restricted sub-cellular localisations, a quality fundamental to their ability to integrate specific subsets of Ca2+ signals that are in turn linked to particular cellular pathways.
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