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. 2017 Jun 30;292(34):13970–13985. doi: 10.1074/jbc.M117.778134

Figure 9.

Figure 9.

CHGA promoter haplotype 2 confers higher expression of CHGA. A, schematic representation of the CHGA promoter haplotype-cDNA constructs. The CHGA cDNA was subcloned in-frame into the Hap1–GLuc, Hap2–GLuc, and Hap5–GLuc constructs replacing the Gaussia luciferase cDNA. B and C, in vitro expression of CHGA under influence of the CHGA promoter haplotypes. N2a cells were transfected with Hap1-CHGA, Hap2–CHGA, and Hap5–CHGA constructs and Western blotting of total proteins was carried out to probe for CHGA and β-Actin. Fold-difference (CHGA/β-Actin) is indicated. D, plasma CHGA levels in Hap1/Hap1 subjects (shown as Hap1,1), Hap1/Hap2 individuals (Hap1,2), and Hap2/Hap2 subjects (Hap2,2) as measured by ELISA. E, plasma CHGA levels of Hap2/Hap2 and Hap2/Hap5 individuals. F and G, CHGA promoter haplotypes transfected in diploid combinations. IMR-32 cells were transfected with 1 μg each of CHGA Hap1–GLuc or Hap2–GLuc or 500 ng each of Hap1–GLuc and Hap2–GLuc or 1 μg each of CHGA Hap2–GLuc or Hap5–GLuc or 500 ng each of Hap2–GLuc and Hap5–GLuc constructs. Results are mean ± S.E. of triplicate values of Gaussia luciferase/β-Gal. *, p < 0.05 as compared with Hap1–GLuc construct (F). #, p < 0.05 and ***, p < 0.001 as compared with Hap2 + Hap5 condition and Hap5–GLuc construct, respectively (G).