Figure 1. Expression of synaptic UPS reporters in neurons. (A) Schematic map of DNA constructs for expressing fluorescent UPS reporters. RFP or GFPu is added to the C terminus of PSD95 or SNAP25 in an adenoviral vector to express fusion proteins under the control of the human synapsin 1 promoter. (B) Western blots showing the expression of these fusion proteins in HEK293 cells after adenoviral infection. (C) Expression of fluorescent UPS reporters in primary cultured cortical neurons after infection with adenoviral SNAP25-RFP vector. SNAP25-RFP was expressed in cultured primary neurons (NeuN-positive) but not in cultured glial cells (GFAP-positive). (D) In the mouse brain striatum injected with adenoviral SNAP25-RFP reporter, SNAP25-RFP was also expressed in neurons, but not glial cells. Immunostaining for GFAP or NeuN (green), nuclear staining (blue) by Hoechst, and SNAP25-RFP (red) were revealed by fluorescent microscopy. Bars, 5 μm (C) and 10 μm (D).
Figure 1.

Expression of synaptic UPS reporters in neurons. (A) Schematic map of DNA constructs for expressing fluorescent UPS reporters. RFP or GFPu is added to the C terminus of PSD95 or SNAP25 in an adenoviral vector to express fusion proteins under the control of the human synapsin 1 promoter. (B) Western blots showing the expression of these fusion proteins in HEK293 cells after adenoviral infection. (C) Expression of fluorescent UPS reporters in primary cultured cortical neurons after infection with adenoviral SNAP25-RFP vector. SNAP25-RFP was expressed in cultured primary neurons (NeuN-positive) but not in cultured glial cells (GFAP-positive). (D) In the mouse brain striatum injected with adenoviral SNAP25-RFP reporter, SNAP25-RFP was also expressed in neurons, but not glial cells. Immunostaining for GFAP or NeuN (green), nuclear staining (blue) by Hoechst, and SNAP25-RFP (red) were revealed by fluorescent microscopy. Bars, 5 μm (C) and 10 μm (D).

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