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. 2010 Aug 4;30(31):10472–10483. doi: 10.1523/JNEUROSCI.0257-10.2010

Figure 3.

Figure 3.

FS stress induces tyrosine phosphorylation of SIRPα through activation of SFKs. A, WT or Fyn-deficient (Fyn KO) mice were subjected to FS stress (FS+) or control treatment (FS−) for 10 min, after which homogenates of the hippocampus were immediately prepared and subjected to immunoblot analysis with anti-pSIRPα, pAbs to SIRPα, or a mAb to Fyn. The ratio of the intensity of the pSIRPα band to that of the SIRPα band is presented as fold increase relative to the WT control (FS−) value; data are means ± SEM. for a total of eight WT and nine Fyn-deficient mice for each condition in four independent experiments. *p < 0.05; **p < 0.01, Student's t test. B, Hippocampal homogenates of mice with forebrain-specific deficiency of TrkB (TrkBlox/lox/CaMKII-Cre) or control (TrkBlox/lox) animals treated as in A were subjected to immunoblot analysis with anti-pSIRPα or pAbs to SIRPα. The homogenates were also subjected to immunoprecipitation with pAbs to pan-Trk, and the resulting precipitates were subjected to immunoblot analysis with a mAb to TrkB. The ratio of the intensity of the pSIRPα band to that of the SIRPα band is presented as fold increase relative to the TrkBlox/lox control (FS−) value; data are means ± SEM. for a total of three TrkBlox/lox and four TrkBlox/lox/CaMKII-Cre mice for each condition in three independent experiments. C, Hippocampal homogenates prepared from WT mice treated as in A were subjected to immunoblot analysis with pAbs to the Tyr416-phosphorylated form of c-Src (pY416; left). The homogenates were also subjected to immunoprecipitation with mAbs to Src (middle) or to Fyn (right), and the resulting precipitates were subjected to immunoblot analysis with pAbs to Tyr416-phosphorylated Src or with mAbs to Src or to Fyn. The intensity of the pY416 band (left) and the ratios of the intensity of this band to that of the Src or Fyn bands (middle and right) are presented as fold increase relative to the control (FS−) value; data are means ± SEM. from a total of four mice for each condition in two independent experiments. *p < 0.05, Student's t test. IP, Immunoprecipitation.