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However, when the mice were subject to cold challenge, axon outgrowth response of sympathetic neurons in the celiac ganglia was significantly inhibited by 1-NaPP1, assessedsed by anti-STMN2 immunostaining (Fig. 5C and 5D).
To get insight into the signaling mechanisms that allow the orientation and the steady neurite outgrowth response of the neuronal like cells on the line pattern, we explored if there are global differences in signaling activities in response to ECM topography.
Given the high degree of homology between NgR1 and NgR2 we hypothesized that the lack of a robust phenotype in the neurite outgrowth response to MAG from neurons deficient for either NgR1 or NgR2 was attributed to functional redundancy between the two receptor constituents.
These differences in MDA MB 231 and MCF7 signalling networks are likely the basis for the contrasting outgrowth response noted between the two breast cancer cell lines.
In contrast, the defective PI3K signaling cascade and its associated mTORC2 activity were largely rescued together with the NGF-induced neurite outgrowth response.
In the PC12-27/p75NTR cells, on the other hand, the shape was not changed much, however the NGF-induced neurite outgrowth response was evident in terms of both number of neurites sprouted per cell and average neurite length (Fig. 7E).
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These data suggest that Egr1 is directly required in the sAPPs-induced axon outgrowth and not in dendrite outgrowth responses.
We find that when cells are challenged with a highly defined anisotropic, nanotopographic laminin substrate, distinct neurite outgrowth responses occur in comparison with the classic, isotropic 2D environment.
We then used differentiated N1E-115 cells as a model system to compare the neurite outgrowth responses on classic 2D, laminin-coated coverslip (plain substrate) versus laminin that is presented on these line patterns (line substrate).
To evaluate the neurite outgrowth responses, we stained the microtubule cytoskeleton and the nuclei of the cells at different time points after plating and used automated image analysis to measure neurite length and orientation on the plain and line substrates (Fig. S1B).
Noteworthy, this genetically imprinted region (Seitz et al, 2004) modulates neuronal responses with some miRNAs shown to have a crucial role in dendritic outgrowth in response to the environment (Khudayberdiev et al, 2009).
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