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. 2024 Oct 11;27(11):111165. doi: 10.1016/j.isci.2024.111165

Figure 3.

Figure 3

Compound displaying dual inhibitory activity against AIMP2 and α-synuclein aggregation without estrogenic activity

(A) Trypan blue exclusion cell viability assessment conducted in SH-SY5Y cells transfected with Myc-AIMP2 or β-gal control (72 h) and treated with indicated compounds for AIMP2 and/or α-synuclein inhibitory activity (10 μM, 48 h) (n = 6 per group).

(B) Quantification of estrogen responsive gene, EBAG9 messenger RNA levels in the human breast tumor cell line, MCF7 treated with the indicated compounds (10 μM, 48 h) (n = 5 separate experiments per group). GAPDH served as internal loading control for normalization.

(C) Dot blot assessment of biotin-conjugated SG13-136 (SG13-158: 0, 2, 4, 8, 16, and 32 nmol) binding to recombinant α-synuclein (αSyn) or/and AIMP2 (1 μg). SG13-136 binding to recombinant GST, Tau, and ZNF746 (1 μg) was also monitored to determine the specificity of SG13-136 binding to α-synuclein and AIMP2.

(D) Binding curve depicting increasing doses of biotin-conjugated SG13-136 for each recombinant protein in the dot blot assay. Dissociation constant (Kd) values of biotin-conjugated SG13-136 binding to α-synuclein and AIMP2 are provided. Data in all panels represent mean ± standard error of the mean. ∗p < 0.05 and ∗∗∗p < 0.001, determined by one-way (B) or two-way (A) analysis of variance (ANOVA) followed by Tukey’s post hoc analysis. n.s., non-significant.