Fig. 5.

SMAD3 and SMAD4 as primary targets for LAT-derived miRNAs. (A) HEK293TqRT-PCR analysis of SMAD3 and SMAD4 in HEK293T cells transfected with pCDH-miR-H2, pCDH-mir-H3, pCDH-miR-H4, or pcDNA3-LAT against cells transfected with empty pCDH or pcDNA3(-) as control vectors. Relative levels of gene expression were normalized to B2M mRNA level. ^**p < 0.001; Luciferase activity assay for direct targeting of the 3’-UTR of (B) SMAD3 and (C) SMAD4 by miR-H2, miR-H3, mir-H4, or LAT. Partial segments of SMAD3 and SMAD4 were fused with a luciferase reporter and co-transfected into HEK293T cells with the recombinant plasmid containing miR-H2, -H3, -H4, LAT, or empty pCDH. Luciferase activity was measured 48 h post transfection. Levels of Renilla luciferase activity were normalized to firefly luciferase activity as an internal control. ^**p < 0.0001