2 and ErbB3, it would allow us to express exogenous GM ErbB4 receptor. Second, PC12 cells developed, reported to bring people to express ErbB4 was to differentiate and to undergo neuritogenesis upon treatment with recombinant NRG1, what motivates us, k nnte A Ph Phenotype, quantified using automated microscopy and analysis, k nnten VX-745 VX745 offer picture.Schliemann has Lich NGF induces differentiation has been studied extensively in PC12 cells, and compare a lot of help on the downstream signaling pathways, which motivated the selectivity t of all connections, which we identify h Tte known. To create a system for chemical genetic study, we have a stable PC12 cell line that coexpressed controlled green fluorescent protein and the human ErbB4 isoform Cyt2 JMA and a cell line The PC12-GFP expressed from F Stable GFP lacking, but ErbB4.
The ErbB4 PC12 cells were examined by GFP fluorescence imaging for the effects of NGF and NRG1 on neurite outgrowth. As expected, both the GFP and ErbB4 PC12 PC12 cell lines GFP morphological Ver Changes, the functions of neuronal differentiation with neurite outgrowth when treated with NGF. at low mag AREA u Erte the distribution of GFP cooperation throughout Zellk body and neurite projections uniform, and the fluorescent image of F is reliably, precious metals, is the whole Zellk body and processes listed. NRG1 stimulation of PC12 cells, CFP ErbB4, but not GFP PC12 went for 2 days Born in neurite outgrowth significantly Similar to the effects of NGF.
This result shows that, when PC12 cells express ErbB3, which can also bind NRG1 and heterodimers with other erbB family members, the presence of the ErbB4 receptor for NRG1 signals required to stimulate the neuritogenesis. In addition have to stimulate the treatment of EGF to neurite outgrowth in both cell lines, suggesting that, although EGFR activation can be activated, other ErbB receptors by heterodimerization, not in a position to trigger neuritogenesis in the presence or absence of ErbB4ErbB4 is known that the downstream effectors of canals len neurotrophic factor, such as mitogen activated protein kinase activate. To ensure that GFP activated PC12 cells ErbB4 components of the MAPK pathway in response to NRG1, we examined the F Ability of NGF and NRG1 treatment to activate the MAPK pathway, such as by the phosphorylation of extracellular Ren kinase of the measured signal .
In both cases, The GFP-PC12 and PC12 cell lines GFP ErbB4 induces NGF rapid phosphorylation of ERK1 / 2, which tt than 5 minutes after treatment, their H Hepunkt. In contrast, a strong NRG1 phosphorylation of ERK1 / 2 only induced in PC12 cells GFP ErbB4. In addition, it was reported that NRG1 ht obtained The release of intracellular Ren Cathedral Ne of ErbB4, ErbB4 ICD in the neural precursor Shore cells. However, we did not observe this effect in PC12 cells. We also found that an inhibitor γ secretase cleavage of the ErbB4 NRG1 inhibits not inhibit neurite outgrowth in PC12 cells induced ErbB4. After the Best Confirmation that stimulation of PC12 cells with GFP ErbB4 NRG1 k Nnte induce neuritogenesis, we sought next to determine whether the growth of Ph Genotype suitable for use with automated image processing and image analysis was. The L Soluble GFP expression in PC12 cells erm Glichte the use of automated microscopy to acquire