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. 2016 Mar 24;6:23464. doi: 10.1038/srep23464

Figure 3. NS5A expression in HEK293 cells and Huh-7 cells.

Figure 3

(A) GFP-NS5A is localized to ER membranes (inset 1 and bracket, inset 3) and induces formation of lipid droplets (inset 2). GFP-NS5A frequently fragments and encircles mitochondria (arrow, inset 3). In this experiment, HEK293 cells were transfected with GFP-NS5A for 18 hours before fixation and staining with mtHSP70 (red). (B) TdTomato-NS5A (red) transfected into Huh-7 cells and stained with lipid droplet marker bodipy 493/503 (green). NS5A encircles lipid droplets (arrows). (C) EM study of the effect of NS5A in ER-mitochondria tethering in HEK293 cells transfected with Myc-NS5A or pMyc vector. Extensive ER-tethered mitochondria was observed in Myc-NS5A transfected HEK293 cells but not in Myc vector transfected cells. Scale bars = 1 μm. Abbreviations: M, mitochondria; ER, endoplasmic reticulum. (D) Quantification of ER-mitochondria tethering in NS5A-transfected HEK293 cells from EM images. The percentage of ER-tethered mitochondria is determined by the length of the mitochondria perimeter apposed to ER tubules in the EM images. N = 60; error bar = S.E.M.