Skip to main content
. 2010 Jul 5;19(18):3634–3641. doi: 10.1093/hmg/ddq279

Figure 4.

Figure 4.

L253P β-III spectrin does not interact with Arp1 and interferes with protein trafficking. (A) Neuro2a cells cotransfected with EAAT4 and either myc-tagged WT or L253P β-III spectrin. Cells immunostained using anti-c-myc antibody (red), anti-EAAT4 antibody (green) and nucleus stained with DAPI (blue). (B) Cells transfected with either myc-tagged WT or L253P β-III spectrin and 24 h after transfection incubated at 37 or 25°C for a further 12 h. Cells immunostained using anti-c-myc antibody (red). Western blot analysis of cell homogenates probed with anti-c-myc antibody. (C) Cells expressing myc-tagged L253P β-III spectrin and EAAT4 incubated at 25°C for a further 12 h. Cells immunostained using anti-c-myc antibody (red) and anti-EAAT4 antibody (green). (D) BiFC assay using cells transfected with Arp1 fused to the N-terminal half of YFP (YN-Arp1) and the actin-binding region of β-III spectrin, with or without the L253P substitution, fused to the C-terminal half of YFP (YC-β-III). Twenty-four hours after transfection cells were incubated at 37 or 25°C for a further 12 h. Western blot analysis of cell homogenates probed with anti-GFP antibody. All images are representative of three independent experiments (scale bar, 10 µm).