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. 2015 Mar 25;200(1):221–235. doi: 10.1534/genetics.115.175596

Figure 2.

Figure 2

Gimap3 localizes to the endoplasmic reticulum. (A) Sedimentation profile of mouse postnuclear spleen lysate through a discontinuous Histodenz gradient. Fractions were separated by SDS–PAGE followed by immunoblotting with the indicated antibodies. Canx is an ER chaperone. Sdha and Porin are mitochondrial proteins localized in the matrix and outer membrane respectively. (B) A graphical illustration of the Gimap3 constructs used in microscopy. (C) Confocal images of COS-7 cells cotransfected with a chimeric RFP–Gimap3 and ER marker GFP–Sec61B. Below, line scans of the red and green channels. (D) Confocal images and inset magnifications of COS-7 cells cotransfected with the chimeric RFP–Gimap3 and the mitochondrial marker GFP–omp25. (E) COS-7 cells transfected with a construct containing only the C-terminal transmembrane domain of Gimap3 fused to YFP. (F) COS-7 cells transfected with a construct containing the full-length Gimap3 fused to the C terminus of YFP. (G) COS-7 cells cotransfected with RFP–Gimap3 and a chimeric YFP–Gimap3 missing the 32 C-terminal amino acids replaced with the omp25 sequence motif for mitochondrial localization. Scale bars, 10 µm.