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Figure 3 | Molecular Brain

Figure 3

From: Dopamine D1 receptor-mediated NMDA receptor insertion depends on Fyn but not Src kinase pathway in prefrontal cortical neurons

Figure 3

Efficiency of Src and Fyn knockdown in cultured PFC neurons. (A) 11 DIV PFC neurons were transfected for 48 h with Fyn siRNA and stained with anti-Fyn and PSD95. Fyn, but not PSD95, decreased dramatically after siRNA transfection. Scale bar = 10 μm. (B) 11 DIV PFC neurons were transfected for 48 h with Src siRNA and stained with anti-Src and PSD95. Similarly, Src, but not PSD95, was significantly reduced in transfected neurons. Scale bar = 10 μm. (C and D) Quantifications of Fyn and Src immunofluorescence in dendrites suggest the effective knockdown of both Fyn and Src kinases in the cultured PFC neurons (n = 30, ** p < 0.01, *** p < 0.001). (E-H) Total protein levels of Fyn and Src were determined by Western blotting after Fyn and Src siRNA transfection in cultured PFC neurons. The blots were stripped and reprobed with antitubulin as loading control and the siRNA knockdown specificities were demonstrated by re-probing the Src and Fyn, respectively. Integrated intensity analysis showed significant decreases in both Fyn and Src proteins after knockdown (n = 3, ** p < 0.01). Control protein levels of Fyn and Src were set at 100% after being normalized to loading control tubulin.

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