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Strong reduction of CENP-H resulted in a slightly reduced CENP-C level at the kinetochores and normal localisation of hBubR1, indicating a functional mitotic checkpoint at the hBubR1 protein level.
In stage 16 gho mutant embryos, in addition to a variable but occasionally normal localisation at the apico-lateral membrane, the Crb signal accumulates within the cell (Figure 5D).
The cell-cell contact protein β-catenin also showed normal localisation around the plasma membrane of the cardiomyocytes (Hirschy et al., 2006) coupled with some nuclear signal.
Whereas E-cadherin, P-cadherin and DSP were completely displaced from cell cell junctions in SJG15 cells, they exhibited a largely normal localisation in other cancer cells (Supplementary Fig. 5).
In comparison to the normal localisation of wild-type CaBP5 (Fig. 1F) both the CaBP5CaBP7TMD and CaBP5CaBP8TMD fusion constructs (Fig. 4A and B) co-localised extensively with the TGN marker ts045 VSVG-GFP.
However, normal localisation of Irgm2 and Irgm3 to the T. gondii PVM was restored in cells transfected with the GKS proteins Irga6, Irgb6 and Irgd in addition to the three GMS proteins and when cells transfected with either Irgm2 or Irgm3 alone were co-induced with IFNγ (data not shown).
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Both proteins show normal localisations in small clones with defective epithelial organisation, whereas aPKC and Par-6 levels are already elevated in small clones that still retain a normal shape (Fig. 4H-K).
However, when MIF-induced gs3T3-Irga6 cells were simultaneously induced with IFNγ, normal ER localisation of Irga6 was restored and the protein could also accumulate normally at the PVM.
The normal cytoplasmic localisation of chicken Mx may influence its antiviral capacity.
Group 3 mutant channels were non-functional, due to an almost complete lack of protein at the plasma membrane (T187I, W1421X, K1578fs/52, R1623X) or a probable gating/permeation defect with normal surface localisation (R878C, G1408R).
Collectively these data show that fluorescent protein tagging of CaBP7 or CaBP8 either N- or C-terminally with mCherry or EYFP has no impact on normal protein localisation as assessed by comparison and colocalisation with endogenous CaBP7.
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