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. 2016 Jun 21;291(32):16462–16476. doi: 10.1074/jbc.M116.730440

FIGURE 6.

FIGURE 6.

PKCϵ is essential for the expression of PSD-95 and synaptophysin. Primary human neurons were transfected with empty vector/scrambled siRNA (C), PKCϵ siRNA (PKCϵ KD), or a PKCϵ overexpression vector (PKCϵ OE) following the method described under “Experimental Procedures.” Cells were analyzed 72 h after treatment. A, mRNA transcript levels of PKCϵ. B, PSD-95. C, synaptophysin. D, SNAP-25. E, syntaxin-1 in PKCϵ KD and PKCϵ OE neurons. PKCϵ KD suppressed, while PKCϵ OE induced PSD-95 and synaptophysin mRNA transcript. F, agarose gel image showing no effect of scrambled siRNA on PKCϵ mRNA. G, immunoblot showing protein expression of PKCϵ in untreated, PKCϵ siRNA, and scrambled siRNA-treated human neurons. H, immunoblot staining of PKCϵ, PSD-95, synaptophysin, and actin in control, PKCϵ KD, and PKCϵ OE neurons. I–K, graphical representation of protein expression levels of PKCϵ, PSD-95, synaptophysin in control, PKCϵ KD, and PKCϵ OE neurons. Data are represented as the mean ± S.E. of three independent experiments (Student's t test. *, p < 0.05; **, p < 0.05; ***, p < 0.0005).