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. 2022 Jul 7;15:882396. doi: 10.3389/fnmol.2022.882396

FIGURE 2.

FIGURE 2

Allele-specific effects of rs61753730 on FZD6. (A) Effect of rs61753730 mutation on predicted structure of FZD6 protein generated by PyMOL. Upper panel: Naive 3D structure of FZD6 and mutant structure of rs61753730 (Q152E). Bottom panel: Naive structure of FZD6 shows glutamine at position 152, and predicted mutant structure of rs61753730 shows glutamic acid at corresponding position. (B) Effect of rs61753730 mutation on secondary structure of FZD6 at position 152 predicted by NetSurfP-2.0. Box with dashed line in red indicates different structure of WT (C) and mutant SNP (G). (C) Effect of rs61753730 on luciferase activity. HEK293T cells were transfected with rs61753730 C- and G-allele constructs of pGL3-promoter vector. Luciferase activity was measured 48 h later. Data are reported as relative firefly luciferase activity normalized to Renilla luciferase activity and represented as mean ± SEM (n = 3). *p < 0.05. (D) Effect of rs61753730 on mRNA stability of luciferase reporter gene. HEK293T cells were transfected with rs61753730 C- and G-allele constructs of pGL3-promoter vector for 24 h prior to adding actinomycin D (5 μg/ml). The mRNA of luciferase expression was measured at different times (0, 1, and 3 h) and normalized to GAPDH. Data are presented as mean ± SEM (n = 3). *p < 0.05.