Analysis of protein expression and processing in lysates of transfected cells by Western blot.

<p>Cell lysates (30μg total protein) of Expi-HEK293 transfected with plasmids ZO2 (CprME), or ZO3 (NS2B/NS3Pro) or both (ZO2/ZO3) were probed with anti-Zika protein specific polyclonal antibodies (See <a href="http://www.plosntds.org/article/info:doi/10.1371/journal.pntd.0005608#sec002" target="_blank">Material and Methods</a>). Zika virus (ZIKV) infected and uninfected (mock) Vero cell were controls. <b>(A)</b> Anti-E specific antibody detected E protein expression and cleavage in ZO2/ZO3 and ZO2 (lesser amount) cell lysates but not in that of ZO3. E was also detected in ZIKV infected cells but not in the mock control. <b>(B)</b> The prM protein (~32KDa) was detected in ZO2/ZO3 transfected cells as well as in ZIKV infected cells, which also showed a small amount of M (~ 8KDa). <b>(C)</b> NS3Pro (~19KDa) was detected in ZO3 and ZO2/ZO3 transfected cells whereas the full-length protease/helicase NS3 (~69KDa) was detected in ZIKV infected cells. Neither was detected in ZO2 transfected cells or the mock control. <b>(D)</b> NS2B (~14KDa) was detected in ZO2 and ZO2/ZO3 transfected cells and ZIKV infected cells whereas ZO2 and mock transfected cells were negative.