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. 2010 Oct 31;286(3):2155–2170. doi: 10.1074/jbc.M110.188482

FIGURE 5.

FIGURE 5.

A, luciferase assay used to measure putative minimal FTO (FTO1p and FTO2p) and RPGRIP1L (RPGRIP1Lp) promoter activity upon human P200 ((pCMV) P200) or P110 ((pCMV) P110) overexpression or transfection with empty pCMV and in the presence or absence of the putative enhancer carrying the CUX1-binding A (Enh(A)) or C (Enh(C)) alleles. To control for background, extracts from cells transfected with empty pGL3 or pGL3 carrying the putative enhancer (Enh(A)+Enh(C)) in the absence of FTO1p, FTO2p, and RPGRIP1Lp were also assayed for luciferase activity. Transfection with 20 ng of Enh(C):FTO1p, Enh(C):FTO2p, or Enh(C):RPGRIP1Lp pGL3-based plasmids resulted in off-scale fluorescence intensity; only 2 ng of the above plasmids was used in this experiment. B, expression analysis in N41 cells overexpressing p200 or p110. C, expression analysis in primary neuronal cultures treated with leptin and/or cathepsin L inhibitor I. Each bar represents n = 3. Experiments were repeated twice. *, statistically significant. Used only for comparisons of data within close range (p values <0.01). Error bars represent S.D.