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. 2011 Mar 14;6(3):e17545. doi: 10.1371/journal.pone.0017545

Figure 4. Efficiency of BTV-1 VP2 cleavage by C. sonorensis saliva is temperature dependent.

Figure 4

BTV-1 particles (2 µg) were incubated with 1 µg of C. sonorensis saliva proteins at 4°C, 10°C, 15°C, 20°C, 25°C or 37°C for 2 hours in a final volume of 50 µl. Viral proteins were analysed on 10% SDS-PAGE and visualized using silver staining. Lane M shows molecular weight markers. Lane 1 and 8 are purified BTV-1 virus particles incubated at 37°C in the absence of proteases. After incubation at 4°C and 10°C (lane 2 and 3) intact BTV-1 VP2 was still visible, however some cleavage of VP2 had occurred resulting in intermediate cleavage products (ICP). At 15°C, no intact VP2 was visible but still only intermediate cleavage products (ICP) are generated. (lane 4). At higher temperatures, intermediate CP were completely cleaved to final CP (lane 7).