BAC-K297R mutant virus is colocalized with the lysosomes.

<p>iSLK-BAC16 and iSLK-BAC-K297R cells were induced with Dox (3 μg/ml) for 3 days for viral lytic replication. The double-labeled IFA was performed with mouse monoclonal anti-ORF65 and rabbit polyclonal anti-LAMP-1. Alexa-555 conjugated anti-mouse IgG and Alexa-647 conjugated anti-rabbit IgG were used as the respective secondary antibodies. The channels of Alexa-555 and Alexa-647 were defined in green and red, respectively. The colocalization sites were enlarged as shown (A–H). iSLK-BAC16 and iSLK-BAC-K297R cells were induced with Dox and butyrate and lysed by RIPA lysis buffer at indicated time of post-induction. The cell lysates were resolved on 8% SDS-PAGE and analyzed by Western blotting with antibodies against tegument proteins ORF64, ORF45 and lysosome marker LAMP-1, respectively. β-actin serves as a loading control (I). The intensity of each band was quantitated using ImageJ software (J–M).