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Fig. 2 | BMC Research Notes

Fig. 2

From: Expression of dengue virus and Zika virus NS2B-NS3pro constructs alter cellular fatty acids, but co-expression with a Zika virus virus-like particle is detrimental to virus-like particle expression

Fig. 2

Analysis of ZIKV E protein expression from bicistronic constructs. HEK293T/17 cells were transfected with either a ZIKV VLP (ZVLP) or co-transfected with a ZIKV VLP and either a plasmid expressing EGFP, or a plasmid expressing ZIKV NS2B-NS3pro (Zwt), or a plasmid expressing ZIKV NS2B-NS3pro (S135A) (ZMut), or a plasmid expressing DENV NS2B-NS3pro (Dwt), or a plasmid expressing DENV NS2B-NS3pro (S135A) (DMut), or transfected with one of seven bicistronic vectors containing a ZIKV VLP downstream of either EGFP ((EGFP) or ZIKV NS2B-NS3pro (Zwt), ZIKV NS2B-NS3pro (S135A) (ZMut), ZIKV NS3pro (ZNS3), DENV NS2B-NS3pro (Dwt), DENV NS2B-NS3pro (S135A) (DMut) or DENV NS3pro (DNS3) separated by a cell cleavableP2A linker. At day 3 post transfection cells and supernatants were harvested and proteins extracted and separated by SDS-PAGE. (A) Proteins were transferred to membranes which were probed with a pan-flaviviral E protein antibody (HB112), or (C) a rabbit polyclonal anti-Zika virus E protein antibody (Zika E), or a rabbit polyclonal anti-Zika virus NS3 antibody (Zika NS3), or a mouse monoclonal anti-β actin antibody followed by appropriate HRP-conjugated secondary antibodies. Signals were captured by an AZURE C400 chemiluminescent gel imaging system and (B, D) quantitated using ImageJ. Error bars indicate the standard error of the mean from 3 independent biological replicates. Full length uncropped western blots are given in the supplemental materials. P value * < 0.5, ** < 0.01. Bands shown are cropped from full length gels, and full length uncropped western blots are given in the supplemental materials

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BMC Research Notes

ISSN: 1756-0500

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