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. 2014 Mar 24;33(37):4613–4622. doi: 10.1038/onc.2014.66

Figure 1.

Figure 1

The EBV oncogene LMP1 increases endogenous levels of the transcription factor HIF1α in exosomes secreted by NP cells. (a) Purified exosomal fractions from NP69 cells stably expressing LMP1 or the control and AdAH nasopharyngeal cells transiently transfected with LMP1 or a control-expression vector were analyzed by immunoblotting. Total protein lysates of exosome pellets, isolated from supernatant fluids, were normalized to exosome-specific markers HSC70 and flotillin2. (b) Purified exosomes from NP69-LMP1 and 293T cells transiently transfected with HIF1α expression plasmids were subjected to ultrastructural examination and immunogold labeling for HIF1α. Exosomes were stained with HIF1α primary and colloidal gold-labeled secondary antibodies. (c) LMP1-negative AdAH cells were treated with graduated amounts of exosome fractions isolated from LMP1-expressing AdAH cells. After 24 h, western blotting analyses were used to detect LMP1, HIF1α and GAPDH (glyceraldehyde 3-phosphate dehydrogenase) expression in recipient AdAH cells.