10.1371/journal.pone.0143467.g003 Jie Zhu Jie Zhu Binbin Wang Binbin Wang Qiuhong Miao Qiuhong Miao Yonggui Tan Yonggui Tan Chuanfeng Li Chuanfeng Li Zongyan Chen Zongyan Chen Huimin Guo Huimin Guo Guangqing Liu Guangqing Liu The interaction between VPg and eIF4E in RK13 cells based on the mammalian two-hybrid assay. Public Library of Science 2015 novel cap substitute Rabbit hemorrhagic disease virus RHDV translation vpg tissue culture system eukaryotic initiation factor 4 E rabbit hemorrhagic disease eIF 4E rk RHDV replicon 2015-11-23 15:22:07 Figure https://plos.figshare.com/articles/figure/_The_interaction_between_VPg_and_eIF4E_in_RK13_cells_based_on_the_mammalian_two_hybrid_assay_/1610228 <p>RK13 cells (2×10<sup>5</sup> cells per well in a 6-well plate) were co-transfected with the pACT (1 μg) and pBIND (1 μg) plasmids, pACT-VPg (1 μg) and pBIND (1 μg) plasmids, and pACT-VPg (1 μg) and pBIND-eIF4E (1 μg) plasmids, respectively. RK13 cells co-transfected with the pACT-MyoD (1 μg) and pBIND-ID (1 μg) plasmids served as positive controls [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0143467#pone.0143467.ref025" target="_blank">25</a>]. A blank control was also used. In addition, all of the groups were transfected with pGL4.75 (0.2 μg). The cells were lysed at 48 h post-transfection, and Fluc activity was evaluated based on RLUs and normalized according to the results obtained for a co-transfected plasmid encoding the Renilla luciferase. The experiments were conducted in triplicate, and similar results were obtained from three independent experiments. The Student’s <i>t</i> test and ANOVA were applied for the statistical analyses, <i>P</i> <0.01 was considered as extremely significant different (**).</p>