Fig 2: OLFM4 expression was different between septic and sepsis-induced ARDS patients. (A and B) Heatmap and statistical analyses of expression of the ten hub genes in the GSE66890 dataset. Red = upregulated. Blue = downregulated. **p < 0.01. (C) Analyses of ROC curves of critical DEGs in the GSE66890 dataset. ROC curves were generated and the area under the ROC was used to compare the ten genes in the sepsis group and sepsis-related ARDS group. Three DEGs showed AUC >0.7: 0.716 for OLFM4, 0.719 for LCN2, and 0.735 for BPI.

Fig 3: Inhibition of double-strand break repair promotes tubular epithelial cell damage.(A and B) Nanoparticles (50 µg) containing fluorescent ICG dye were injected into mice and monitored 24 hours later using IVIS system in (A) living mice and (B) ex vivo excised organs. (C) Ex vivo organ fluorescence accumulation (n = 3 animals). K, kidney; Lu, lung; Liv, liver; H, heart; Spl, spleen. Data represent mean ± SD. (D–G) Representative histological images from (D) vehicle- or (E and F) cisplatin-treated (15 mg/kg IP) Trip13Stop/Stop (D and E) and Trip13?Stop (F) mice. (G) Tubular epithelial cell damage was scored as a percentage of total tubules counted. Serum markers of AKI were monitored for (H) creatinine and (I) NGAL after 72 hours following treatment with cisplatin (15 mg/kg IP) and NP-mirin or NP-Ctrl (50 mg/kg IP). (J) ?-H2AX– (Ser139) and (K) PCNA-positive nuclei in kidney sections at 72 hours following cisplatin treatment with and without mirin. NP without mirin (50 mg/kg IP) was used as the control solution. (L) Relative gene expression change following treatment with cisplatin and/or mirin. Each of the respective groups was compared with mouse kidney values obtained from control vehicle-treated Trip13Stop/Stop mice. (G–K) n = 5–7 mice/group; (L) n=4 mice/group. *P < 0.05, **P < 0.01 between all organs (C) or indicated groups (G, I, J, and L) or all groups (C) using 1-way ANOVA with Tukey’s post hoc analysis. Significance for creatinine (H) was determined using Kruskal-Wallis nonparametric test with Dunn’s post hoc analysis. Scale bar: 100 µm. + = administration of cisplatin or mirin; Cre - = Trip13Stop/Stop; Cre + = ?Stop.

Fig 4: Trip13 overexpression in the proximal tubules provides protection from renal tubular epithelial cell damage following cisplatin administration.Mice were injected with a single injection of vehicle (20% captisol in saline) or cisplatin (15 mg/kg IP), and kidneys and blood were harvested to detect markers of renal dysfunction. (A) Kidney samples from homozygous floxed stop (Trip13Stop/Stop) and Trip13?Stop mice were harvested from mice treated with either vehicle (-Cis; 20% captisol) or cisplatin (+Cis; 15 mg/kg IP) after 72 hours following the initial injection. Western blot analysis was performed using FLAG and GFP primary antibodies. GAPDH was used as a loading control. Arrows on the right side of the panel indicate protein standard size. (B) Serum creatinine and (C) NGAL was measured in each mouse group. (D) Percentage of tubular damage was determined in each group. (E–H) Representative histological images from (E and G) vehicle- or (F and H) cisplatin-treated (15 mg/kg IP) Trip13Stop/Stop (E and F) and Trip13?Stop (G and H) mice. Tubular epithelial cell damage was scored as a percentage of total tubules counted. — = vehicle; + = cisplatin. *P < 0.05, ****P < 0.0001 between all other groups using 1-way ANOVA with Tukey’s post hoc analysis. n = 4–8 animals per group. Scale bar: 200 µm (E–H).

Fig 5: Effect of EGCG (free and encapsulated) pretreatment on (A) serum KIM-1 level, (B) serum NGAL level, (C) NLPR-3 gene expression level, and (D) caspase-1 gene expression level. Data expressed as mean ± SD (n = 8/group), significant difference vs. arespective control, brespective cisplatin group, and crespective free EGCG group each at p ? .05.