Тема: puma shoes for women
Proteins extracted from three EGFRvIII-expressing puma slides GBM xenografts, as named in panel b, were subjected to immunoprecipitation/western blotting to determine the extent to which EGFRvIII interacts with co-expressed proapoptotic proteins. An EGFR antibody (Ab) was used to immunoprecipitate EGFRvIII. (D) EGFR interacts with PUMA in GBM cell lines. Using proteins purified from three EGFR-expressing GBM cell lines, immunoprecipitation/western blotting was conducted to investigate the interaction between EGFR/EGFRvIII and PUMA.
An EGFR Ab was used to immunoprecipitate EGFR, whereas control IgG was used as negative control for immunoprecipitation. Top panel: immunoprecipitation/ western blotting. Lower panel: western blotting.In light of a previous study [ 25 ] showing that ErbB4 interacts with puma fenty Bcl-2 and the fact that EGFR/EGFRvIII and ErbB4 share structural homology, we examined whether EGFR/EGFRvIII forms a complex with the three proapoptotic proteins that co-express with the receptors.
Collectively, these results indicate that EGFR and fenty by puma EGFRvIII both paradoxically co-express with proapoptotic proteins and specifically interact with PUMA in human GBM.Using a cohort of 101 gliomas and 12 normal brain tissues, we examined the levels of EGFR/EGFRvIII and PUMA via IHC. The results indicate a significant positive correlation between expression levels of EGFR/EGFRvIII and PUMA (p=0.039).
2b in which a GBM (top panel) shows positive staining for both EGFR/EGFRvIII and PUMA and in contrast, a grade II glioma puma thunder spectra (bottom panel) stains negatively for both proteins. The antibody used in the EGFR/EGFRvIII IHC recognizes a C-terminal epitope present in both EGFR and EGFRvIII receptors, and consecutive tumor sections were used in these studies. The high degree of EGFR/EGFRvIII-PUMA co-expression in primary GBM specimens is consistent with the observations in Fig.
As expected, untreated cells showed no evidence of apoptosis (data not shown). The western blotting ( Fig. 3b ) indicated that PUMA expression was specifically and efficiently down-regulated by the PUMA-specific siRNA.Effects of PUMA expression down-regulation and increased EGFRvIII expression on the induction of apoptosis in GBM cells. Extent of apoptosis is computed as: (# of apoptotic nuclei) / (total # of nuclei). In both panels, 250-300 puma football boots cells were examined in each experiment and three independent experiments were conducted to derive means and standard deviations.
(C) Increased EGFRvIII expression confers resistance of GBM cells to apoptosis. U87MG-vector and U87MG-EGFRvIII stable transfectant cells were exposed to 100 ng/ml anisomycin for 48 hrs and subjected to the TUNEL assay to determine the extent of apoptosis. Generation and characterization of the isogenic cell lines were described previously [ 9 ]. (D) Western blotting indicates that U87MG-vector cells express a very low level of endogenous EGFR (higher molecular weight; 175 [=http://www.simplypotterheads.com/puma-fenty-p-654.html]http://www.simplypotterheads.com/images/lose/puma fenty-286dtd.jpg[/] kD) whereas U87MG-EGFRvIII cells contain significant EGFRvIII (140 kD) expression.