Journal: Advanced science (Weinheim, Baden-Wurttemberg, Germany)

Article Title: Small Extracellular Vesicles Orchestrate Cisplatin-Induced Ototoxicity: Potential Biomarker and Targets Discovery.

doi: 10.1002/advs.202502627

Figure Lengend Snippet: Figure 5. Proteome analysis and validation of the Cis-sEV and Ctrl-sEV groups. A) PCA analysis of proteins in the Cis-sEV and Ctrl-sEV groups. B) A Venn diagram indicating the unique and common proteins between Cis-sEV and Ctrl-sEV. C) A Venn diagram depicting the overlapping proteins among the total 694 proteins found in the sEVs and the top 100 sEV proteins from the ExoCarta and Vesiclepedia databases, respectively. D) Volcano plot analysis shows the 240 differentially expressed proteins, including 90 upregulated proteins and 150 downregulated proteins in the Cis-sEV group. E) GO analysis of the cellular components for the 90 differentially enriched expression proteins. The color bar represents the relative Q value, and a Q value <0.05 indicates significant differences. F) Heat map analysis for the eight proteins associated with the apoptosis, autophagy, proteostasis, and inflammasome pathways. The color bar represents the relative expression level. G) Expression verification of CLTC, CCT2, ANXA6, HSPA8, and HNRNPK in the Ctrl-sEV and Cis-sEV derived from cochlear explant, normalized according to BCA protein quantification results. The experiment was repeated two

Article Snippet: Antibodies against CLTC (Abcam, ab21679, 1:1000), CCT2 (Abcam, ab92746, 1:1000), ANXA6 (Proteintech, 12542-1-AP, 1:1000), HSPA8 (Santa Cruz Technology, sc-7298, 1:200), and HNRNPK (Santa Cruz Technology, sc28380, 1:200) were also used in this study.

Techniques: Biomarker Discovery, Expressing, Derivative Assay

Journal: Advanced science (Weinheim, Baden-Wurttemberg, Germany)

Article Title: Small Extracellular Vesicles Orchestrate Cisplatin-Induced Ototoxicity: Potential Biomarker and Targets Discovery.

doi: 10.1002/advs.202502627

Figure Lengend Snippet: Figure 6. Verification of protein expressions in Cis-sEV using both in vitro and in vivo ototoxicity models. A) Viability of HEI-OC1 cells treated with various concentrations of cisplatin for 24 h. n = 3. All results showed as the mean ± SD, with “**” for p < 0.01, and “***” for p < 0.001. B) Western blot analysis in the HEI-OC1 model confirmed the expression of damage-associated molecules after 15 μM cisplatin treatment for 48 h. 𝛽-Actin serves as the reference protein. C,D) TEM analysis of the morphology of Cis-sEV and Ctrl-sEV in the HEI-OC1 damage model. Scale bar = 100 nm. E) Both positive and negative markers (Alix, CD63, Calnexin) were analyzed by western blotting for Ctrl-sEV and Cis-sEV in the cisplatin-induced HEI-OC1 model, normalized according to BCA protein quantification results. F,G) NTA analysis of Ctrl-sEV and Cis-sEV in the HEI-OC1 damage model. H) Validation of CLTC, CCT2, ANXA6, and HSPA8 expression in sEV within the HEI-OC1 damage model, normalized according to BCA protein quantification results. The experiment was performed independently and repeated three times. I) A flowchart depicts the mouse model for cisplatin-induced ototoxicity. For a week, the 4–5-week-old C57BL/6J mice received daily i.p. injections of either cisplatin (3 mg kg−1 day−1) or an equivalent amount of saline. J) The analysis of the ABR threshold was performed on mice exposed to either cisplatin or saline for 7 days. Saline group: n = 3; Cis group: n = 5. The “n’” indicates the number of mice included in the statistics analysis. Results are presented as mean ± SD, with p < 0.01 indicated as “**” and p < 0.001 indicated as “***.” K) Representative images of HCs in the mouse cochlea treated with saline or cisplatin, including the apical, middle, and basal turns. Myosin7a

Article Snippet: Antibodies against CLTC (Abcam, ab21679, 1:1000), CCT2 (Abcam, ab92746, 1:1000), ANXA6 (Proteintech, 12542-1-AP, 1:1000), HSPA8 (Santa Cruz Technology, sc-7298, 1:200), and HNRNPK (Santa Cruz Technology, sc28380, 1:200) were also used in this study.

Techniques: In Vitro, In Vivo, Western Blot, Expressing, Biomarker Discovery, Saline