It is difficult to track virus-coded proteins simultaneously if they localize to multiple subcellular organelles. Here, we present a protocol for the simultaneous detection of dual subcellular localized dengue virus protease by co-transfection. We describe steps for cell seeding, co-transfection with mitochondria targeted red fluorescent protein, cell fixation, permeabilization, and staining of transfected cells with Hoechst stain. Further, we describe how to generate fluorescent intensity profiles using the NIS-Elements software. We then detail procedures for subcellular fractionation followed by western blotting. For complete details on the use and execution of this protocol, please refer to Gandhi et al..
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| http://dx.doi.org/10.1016/j.xpro.2023.102697 | DOI Listing |
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