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. 2017 Oct 23;37(6):756–767. doi: 10.1038/onc.2017.365

Figure 7.

Figure 7

NTSR2 phosphorylation in B-CLL and recruitment of Giα proteins. (a) Representative western blot analysis of Src phosphorylation in MEC-1 cells overexpressing NTSR2 (pCMV6-NTSR2) in the presence or absence of BDNF (100 ng/ml), pertussis toxin (PTX, 200 ng/ml) or K252a (100 nM) for 24 h. (b) Ratio of phosphorylated Src vs pan-Src protein, normalized against actin. Values are means±s.e.m. of three independent experiments in a.u. (c, d). After immunoprecipitation (IP) of NTSR2 from MEC-1 cells overexpressing NTSR2 (pCMV6-NTSR2) or not (EV), or from B-CLL cells in the presence or absence of BDNF (100 ng/ml) or NTS (40 μM/ml), the immunocomplexes were immunoblotted (IB) with anti-Giα1/2 antibodies. (e, f) After immunoprecipitation (IP) of anti-pan-phosphoprotein was performed on MEC-1 cells overexpressing NTSR2 and B-CLL cells in the presence or absence of BDNF (100 ng/ml) or SR142948A (67 μM), the phosphorylation of NTSR2 was detected by immunoblotting (IB) with anti-NTSR2 antibodies. (g) Apoptotic ratio, assessed by cell death ELISA, in B-CLL in the presence or absence of SR142948A (67 μM) for 24 h. Values are mean ratios of apoptotic cells (±s.e.m.) of three independent experiments from different patients (n=3). Significant P-values are indicated in the graphs *P<0.05, ***P<0.001.