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. 2016 Mar 16;291(20):10747–10758. doi: 10.1074/jbc.M116.722272

FIGURE 3.

FIGURE 3.

Involvement of pro-NGF and p75NTR in LDLR regulation in hepatocyte cells experiments using Huh7 hepatocytes (A–D) and primary mouse hepatocytes (E). Values are given mean ± S.E. A, p75NTR is expressed by Huh7 cells as shown by immunoblotting (75 kDa). The levels are not altered by 5 ng/ml pro-NGF. β-Actin was used as a loading control. B, quantitative PCR. Huh7 cells were stimulated with 5 ng/ml pro-NGF for 6 h that increased the expression of LDLR mRNA. n = 4. *, p < 0.05 for pro-NGF versus C. C, LDLR promoter assay. Cells were transfected with the wild-type LDLR promoter and treated with 5 ng/ml pro-NGF for 24 h. The ratio of firefly versus Renilla luciferase activity is given. n = 4. *, p < 0.05 for pro-NGF versus C. D, cells were stimulated with pro-NGF for 24 h, and LDLR levels were determined. Left, immunoblot. Right, quantification. n = 3. **, p < 0.01 for pro-NGF versus untreated control. E, primary hepatocytes were prepared from wild-type and p75NTR gene-deleted mice and cultured as described under “Experimental Procedures.” Cells were stimulated with 5 ng/ml pro-NGF for 24 h followed by immunoblotting. Left, immunoblots. Right, quantification. n = 3. **, p < 0.01 for pro-NGF versus C in wild-type cells.