Shedding of EMVs containing viral protein following Timer-CVB3 infection of differentiated cells.

<p><i>(A–C)</i> “Fluorescent timer” protein changed from green to red over the span of 6 hours in differentiated NPSCs infected with Timer-CVB3. <i>(D)</i>, <i>(E)</i> Digital magnification of <i>(B)</i> and <i>(C)</i>, respectively, revealed the shedding of EMVs (white arrows) some containing matured “fluorescent timer” protein (pink arrow) between 48 and 53 hours post infection. <i>(F–I)</i> Abundant EMVs containing matured “fluorescent timer” protein were observed in differentiated C2C12 myoblast cells infected with Timer-CVB3. <i>(J)</i> Digital magnification demonstrated the presence of EMVs containing matured “fluorescent timer” protein (pink arrow) near recently-infected cells in green at 3 days PI. <i>(K)</i> By 7 days PI, EMVs containing matured “fluorescent timer” protein (pink arrow) were observed near cells with signs of cytopathic effects.