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. 2019 Nov 23;8(12):1502. doi: 10.3390/cells8121502

Figure 1.

Figure 1

SOD1G93A interfered with the nucleic localization of SOD1WT. (a) SOD1WT-GFP was normally localized in the nuclei of WT primary cultured neurons (first layer panel), whereas, SOD1G93A-GFP failed to enter the nuclei of same neurons (second layer panel). SOD1WT-GFP failed to enter the nuclei of SOD1G93A genotype neurons (third layer panel), and most of the SOD1G93A-GFP was localized in the cytoplasm of SOD1G93A genotype neurons (fourth layer panel). (scale bar is 50 μm) (b) Statistical analysis on the localization of SOD1WT and G93A-GFP in primary cultured WT and SOD1G93A genotype neurons (three time trials). Dark gray: Localization of cytoplasm and nuclei; weak gray: Nucleic localization only. (n = 150 for SOD1WT-GFP expressing WT neurons, 143 for SOD1G93A-GFP expressing WT neurons, 165 for SOD1WT-GFP expressing SOD1G93A background neurons and 159 for SOD1G93A-GFP expressing SOD1G93A background neurons, error bars: Standard deviation). (c) Plasmid constructed for expression of SOD1WT-RFP and SOD1G93A-GFP. IRES was used for co-expression to connect the two genes, SOD1WT-RFP and SOD1G93A-GFP. (d) Three different localization patterns of SOD1 WT-RFP (red) and SOD1G93A-GFP (green) co-expressed in primary cultured neurons. SOD1G93A-GFP was localized in the cytoplasm, whereas, SOD1WT-RFP was detected in the whole cell (upper). In a few cases, SOD1WT-RFP and SOD1G93A-GFP were colocalized in the whole area, but in most cases, cytoplasmic colocalization of SOD1 WT-RFP and SOD1G93A-GFP was detected (down). (scale bar is 10 μm). (e) Statistical analysis on the localization of SOD1WT-RFP and SOD1G93A-GFP in primary cultured neurons (results in triplicates); Left: WT neurons; Right: SOD1G93A genotype neurons. (n = 157 for WT neurons and 175 for SOD1G93A background neurons, error bars: Standard deviation).