Fig. 2.

Analysis of wild-type and chimeric p13/p7 proteins in culture. Expression constructs for wild-type or chimeric p13/p7 proteins were transfected into HEK 293T cells and their intra-cellular localisation and processing by cellular signalases analysed. (A) Top panels – Detection of wild-type p13 and ΔC + p7 protein using 1795 (Alexa-fluor 488 nm secondary). Left column shows p13-specific fluorescence (green channel), middle column shows Alexa-fluor 594 nm-conjugated Concanavalin A, a marker for ER/Golgi membranes (red channel), and right column shows an over-lay incorporating Hoechst staining of nuclei (blue channel). Bottom panels – Detection of chimeric p13/p7 proteins using 1055 (Alexa-fluor 488 nm secondary) (left column), other columns as above. (B) Detection of chimeric proteins via Western-blot (WB) using anti-p7 antibody, 1055, using 10⁶ HEK 293T cells/lane lysed in 200 μl Laemmli buffer. Bands migrating at 7 kDa were identified using lysates containing HCV p7 as controls (black arrow). A higher molecular weight species corresponding to unprocessed ΔC + p7 chimeric protein was also evident (grey arrow). (For interpretation of color mentioned in this figure the reader is referred to the web version of the article.)