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In this set of experiments, reporter cells were transduced with Y2 I-AniI expressing lentivirus, sorted for Y2 I-AniI expression 3 dpt (Fig. 5a), passaged in culture and then sorted based on GFP expression at 15 dpt (Fig. 5b).
In some experiments reporter virus replication declined in a manner suggestive of active viral clearance (Fig. 2C, and see below).
In these experiments, reporter cells were used to generate data for reporter activities, but gene expression and protein assays were accomplished with parental cells.
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As a proof-of-concept experiment, reporter gene expression signatures were generated for 1,186 unique chemical fractions isolated from a natural product library derived from a diverse selection of marine bacteria (c.f., Supplementary Methods)7.
YG designed this study, performed the ChIP experiment, reporter gene analysis, and statistical analysis, and prepared this manuscript.
T-HEp3 cells were stably transfected with plasmids reporting for active ERK as described for Fig. 1B except that in some experiments the reporter was GFP.
Moreover, loss-of-function experiments and reporter assays involving deletion mutants of SOX2 3'-UTR luciferase vectors revealed that miR-126 directly targets the 3'-UTR of SOX2.
In another series of control experiments, a reporter containing the minimal enhancer "Y2Y4" from the sea urchin gene spcyclophillin [14] was used.
Moreover, the interaction of FOXP3Δ2Δ7 to RUNX1, NFAT and NF-kB appeared to be unchanged in co-immunoprecipitation experiments and reporter gene assays, when compared to FOXP3fl and FOXP3Δ2.
Comparing the NF-κB response with magnitude of transgene expression (Figure 4B bottom panel) confirms the enhanced NF-κB activation of Casp8p41 compared to Casp8FL, observed in previous experiments using reporter constructs.
2) Dual luciferase assay data should be supported by experiments demonstrating reporter mRNA integrity and abundance.
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