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

FIGURE 3.

FIGURE 3

SHED-CM suppresses SOD1G85R -induced ER stress and increases HSP70 levels. (A): N2a cells expressing mCherry-SOD1G85R were treated with SHED-CM (30%, 50%, and 70%) for 24 h. Subsequently, immunoblot analysis of Bip and Chop was conducted in relation to ER stress. (B and C): Densitometric quantification of Bip and Chop. ###p < 0.001 vs. WT; ***p < 0.001, **p < 0.01, and *p < 0.05 vs. G85R. (D): Immunoblot analysis of HSP60, HSP70, and HSP90. (E–G): Densitometric quantification of HSP60, HSP70, and HSP90. 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. n.s.: not significant. (H–J): N2a cells expressing mCherry-SOD1G85R were treated with SHED-CM (30%, 50%, and 70%) for 24 h, HSP-related gene mRNA expression of HSP60, HSP70, and HSP90 (Hspd1, Hspa1, and Hsp90aa1, respectively) were analyzed using the SYBR Green-based RT-qPCR assay. The expression levels of mRNAs were normalized to the expression level of β-actin mRNA. Results are presented as means ± SEM of three independent experiments based on the fluorescence intensity of the “mock” (mock = 1). n.s.: not significant.