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. 2014 Nov 12;1(1):ENEURO.0019-14.2014. doi: 10.1523/ENEURO.0019-14.2014

Figure 8.

Figure 8.

Arc increases nuclear Tip60 puncta. A−D, DIV18 hippocampal neurons were transfected with Arc-YFP or YFP as a control, and imaged the next day. Arc-YFP expression was stimulated for 4 h with 50 μM forskolin or DMSO as a control, fixed and stained for endogenous Tip60 (red). Comparing Arc-negative (A) with Arc-positive (B, C) neurons, it was found that overexpression of Arc-YFP induced the formation of endogenous Tip60 nuclear puncta, which associate with Arc-YFP puncta (B, C, insets). 58 ± 4% (n = 24) of endogenous Tip60 puncta were associated with exogenous Arc-YFP spots. Overexpression of YFP alone did not induce the formation of Tip60 hotspots (D). E, DIV18 hippocampal neurons were cotransfected with Arc-YFP (green) and βSpIVΣ5-CFP (blue), treated for 4 h with forskolin, fixed and stained for endogenous Tip60 (red). Overexpression of both Arc and βSpIVΣ5 similarly induced the formation of endogenous Tip60 nuclear puncta, which associated with the Arc- βSpIVΣ5 complex. Scale bars, 2 μm; insets, 0.5 μm. The * indicates immunostaining of endogenous protein.