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A renilla luciferase-expressing control reporter was included.
Briefly, cells were co-transfected with Renilla luciferase and control reporter (pRL-TK) for 48 h.
A control reporter expressing renilla luciferase (RL) driven by EMCV IRES was included.
(A) A control shRNA (Ctrli) or an shRNA targeting eIF4E2 (shE2) were transfected into HEK293 cells with pLRE-fLuc reporter and the control reporter phRL-CMV, together with a plasmid expressing let-7a or a control miRNA.
(A) HEK293T cells were transfected with the firefly luciferase-expressing reporter IFN-β-luc and the renilla luciferase-expressing control reporter TK-renilla, with or without a plasmid expressing the ISG indicated.
(D and E) HeLa cells were transfected with plasmids expressing the shRNA targeting eIF4E2, pLRE-fLuc or pREm-fLuc reporter and a renilla luciferase-expressing control reporter, and a rescue plasmid expressing the wild-type or mutant eIF4E2.
pRLTK Renilla was cotransfected as a control reporter vector.
The internal control reporter construct pRL-SV40 was from Promega.
The control reporter (IL-2 min) does not contain any ORE sequence.
FLUC functions as the control reporter and is expressed independently of the inserted sequence.
Control reporter cells (no co-culture) were stimulated with 50 ng/mL Wnt3a.
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