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. 2022 Feb 3;13:805379. doi: 10.3389/fphar.2022.805379

FIGURE 5.

FIGURE 5

SHED-CM effects the activation of IGF-1 receptor. (A): N2a cells were treated with SHED-CM (50%) for 5, 15, and 30 min. Subsequently, immunoblot analysis of p-IGF-1R was conducted. (B and E): The lysates were analyzed by immunoblotting with antibodies for phosphorylated AKT (p-AKT), phosphorylated ERK (p-ERK), phosphorylated GSK-3β (p-GSK-3β), and β-actin. (C, D, and F): Densitometric quantification of p-AKT, p-ERK, and p-GSK-3β. Results are presented as means ± SEM of three independent experiments based on the fluorescence intensity of the “mock” (mock = 1). ***p < 0.001, **p < 0.01, and *p < 0.05 vs. G85R. (G): N2a cells expressing mCherry-SOD1G85R were treated with SHED-CM (30%, 50%, and 70%) and h-recombinant IGF-2 (same amount as included in SHED-CM) for 24 h. Cell viability was measured using a CCK-8 assay. (H): N2a cells expressing mCherry-SOD1G85R were treated with SHED-CM (30%, 50%, and 70%) and neutralizing anti-IGF-2 antibody for 24 h. Cell viability was measured via a CCK-8 assay. Results are presented as means ± SEM of three independent experiments based on the fluorescence intensity of the “mock” (mock = 100). ###p < 0.001 vs. WT; ***p < 0.001, **p < 0.01, and *p < 0.05 vs. G85R; †p < 0.05.