Fig 2: Activation of ISR in spinal cord after SCI. A–E WB analysis and quantitative analysis of phosphorylation of PERK, HRI, GCN2, and PKR at indicated time points following SCI. n=3 (each group contains 3 mice). B–E One-way ANOVA and Tukey’s multiple comparisons test. *P < 0.05, sham vs. 1 dpi, sham vs. 7 dpi; **P < 0.01, sham vs. 3 dpi, sham vs. 7 dpi; ***P <0.001, sham vs. 1 dpi, sham vs. 3dpi; ****P < 0.0001, sham vs. 3 dpi, sham vs. 7 dpi

Fig 3: Following SCI, ISR signal is activated by autophosphorylation of PERK, GCN2, PKR, and HRI, which phosphorylates Ser51 in eIF2a to attenuate global translation and promote nuclear translocation of ATF4 and stress granule formation. Nevertheless, treatment of ISRIB after SCI alleviates ISR-induced effects to delay neuronal degeneration and improve locomotor function

Fig 4: The analysis of endoplasmic reticulum stress markers GRP78 and CHOP, and PERK-eIF2a-ATF4 pathway in Trp53+/- mouse testes. (A) Representative images of Western blotting for GRP78, CHOP, PERK, eIF2a, phospho-PERK, phospho-eIF2a and ATF4 in Trp53+/- mouse testes of each group. (B) Relative each protein expression compared with control; Values represent the average ± S.E.M from three gels per group. Asterisks indicate a statistically significant difference from control: * P < 0.05, ** P < 0.01, *** P < 0.001 on one-way ANOVA with Duncan's post hoc analysis. (C) Representative images of GRP78 and CHOP-stained in each group. sg, spermatogonia; ps, pachytene spermatocyte; ls, leptotene stage spermatocyte; lc, leydig cell; sc, Sertoli cell; CRIS, chronic restraint-induced stress.

Fig 5: The effect of CM from MDA-MB-231 or PC3 cells on the osteoblastic phenotypic expression in MC3T3-E1 cells transduced with shRrbp1. (A, B) MC3T3-E1 cells transduced with shRrbp1 were cultured in a-MEM+CM-231 or a-MEM+CM-PC3 for 7d, respectively. The protein levels of OCN, BMP2, Smad1/5/9, p-Smad1/5/9, and ß-actin were measured via western blotting analysis. (C) MC3T3-E1 cells transduced with shRrbp1 were cultured in a-MEM+ CM-PC3 and a-MEM+CM-PC3 for 28 d, respectively, followed by ARS staining. Left panel: representative images; right panel: absorbance values at 575 nm of different groups. (D, E) MC3T3-E1 cells transduced with shRrbp1 were cultured in a-MEM+CM-231 or a-MEM+CM-PC3 for 7d. The protein levels of CHOP, IREa, p-IREa, PERK, p-PERK, and ß-actin were measured via western blotting analysis. The data are representative of three independent experiments.