Exosomes secreted from GFP-transfected macrophages contain GFP DNA, RNA, the transcription factor, and expressed protein.

<p>Exosomes from GFP-transfected cells were collected over two days and evaluated for (<b>A</b>): GFP DNA (<b>1</b>) and RNA (<b>2</b>) by PCR analysis. Exosomes secreted from macrophages transfected with empty vector were used as a control (<b>3</b>). (<b>B</b>): Levels of GFP DNA and RNA in exosomes from GFP-transfected macrophages were compared to those from empty vector-transfected macrophages (<b>1</b>), or non-transfected cells (<b>2</b>) by Real-Time PCR analysis. <b>C</b>: expression levels of GFP (30K) in exosomes from GFP-transfected cells (<b>1</b>) or empty vector-transfected macrophages (<b>2</b>) were examined by western blot and compared to the levels of CD63 (53K). Exosomes released from GFP-transfected macrophages contained four orders of magnitude more of GFP DNA and RNA compared to non-transfected macrophages or those transfected with empty vector (<b>A, B</b>); and 6.1 times greater levels of the expressed protein, GFP (<b>C</b>). Exosomes contain substantially higher levels of NF-kb, a transcription factor that involved in GFP <i>p</i>DNA expression, compared to macrophages as demonstrated by western blot (<b>D</b>). AFM images of exosomes revealed differences between: (<b>E</b>) small donut-shaped (empty) exosomes released from non-transfected macrophages, and (<b>F</b>) large spherical (likely filled with the expressed proteins and genetic material) exosomes from catalase-transfected macrophages. The bar: 200 nm.