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. 2010 Oct 4;285(50):39551–39563. doi: 10.1074/jbc.M110.114736

FIGURE 2.

FIGURE 2.

LRF reduction affects the expression of Lpp1 and Atp11c genes. a and b, schematic representation of miRNA localization and putative LRF binding sites in Lpp (a) and ATP11c (b) genes. Gray blocks, LRF binding sites. The dashed line shows the localization of the CpG island spanning from −700 to +500 of the ATP11c gene (b). MEF were transfected with siRNA-LRF and siRNA-Ct. c, after 48 h, total RNA was extracted, and qRT-PCR was performed to quantify ATP11c and Lpp transcripts. d, trans-repression assays in HEK293 cells transfected with LRF-expressing vector along with Lpp promoter luciferase-based vectors in the presence of Renilla vector as internal control. Top, promoter activities were measured 48 h after transfection using the Dual-Luciferase reporter assay system. Bottom, schematic representation of the WT and deleted Lpp promoters cloned in the luciferase reporter vector. e, HEK293 cells were transfected with siRNA-LRF or siRNA-Ct and the Lpp promoter-luciferase vector in the presence of Renilla vector as internal control. All the experiments were performed in triplicate. **, p < 0.01; ***, p < 0.001.