Additional file 1 of Renal clearance of graphene oxide: glomerular filtration or tubular secretion and selective kidney injury association with its lateral dimension
Creators: name: Chen, Wei
name: Wang, Bing
name: Liang, Shanshan
name: Wang, Meng
name: Zheng, Lingna
name: Xu, Si
name: Wang, Jiali
name: Fang, Hao
name: Yang, Pu
name: Feng, Weiyue
Publisher: name: figshare
Year: 2023
Subjects: subject: Biochemistry
subject: Space Science
subject: Medicine
subject: Microbiology
subject: FOS: Biological sciences
subject: Cell Biology
subject: Genetics
subject: Molecular Biology
subject: Physiology
subject: Pharmacology
subject: Chemical Sciences not elsewhere classified
subject: Immunology
subject: FOS: Clinical medicine
subject: Biological Sciences not elsewhere classified
subject: Developmental Biology
subject: Marine Biology
subject: Virology

description: Additional file 1: Table S1. Sequences of primers used for RT-qPCR analysis in mouse kidney. Figure S1. Schematic diagram of Yb3+ labelled on DOTA-PEG functionalized GOs. Figure S2. (a) Dispersion stability of s-GOs and l-GOs in saline solution for 7 days. (b) Free Yb3+ concentration was analyzed by color reaction between xylenol orange (XO) and Yb3+. (c) The labelling stability was calculated as the ratio of the Yb mass on GOs (after subtracting free Yb3+) to the original total mass of Yb on GOs. Figure S3. In vivo fluorescence imaging of the biodistribution of s-GOs and l-GOs in mice and the clearance patterns from the body after i.v. injection of s-GOs/ICG and lGOs/ICG separately (n = 3). Dorsal side (a) and ventral side (b) imaged the in vivo clearance of s-GOs and l-GOs in mice. (c) Ex vivo fluorescence images of s-GO and lGO distribution in the 1) heart; 2) liver; 3) spleen; 4) lung and 5) kidneys. (d) Fluorescence intensity of s-GOs/ICG and l-GOs/ICG in the kidneys at 1, 4, 24, 48 h and day 7 post-injection. Fluorescence images were analyzed using Living Image 4.5.2. The unit of an average radiant efficiency: [p/s/cm2/sr]/[µw/cm2] is defined as the ratio of radiant flux per second in per area and per steradian to the excitation power density. Figure S4. Blood clearance curve in mice after i.v. injected of 5 mg/kg bw s-GOs and lGOs. n = 6. Figure S5. Biodistribution profiles of s-GOs and l-GOs after i.v. injection in mice. n = 6. Figure S6. (a) Typical FTIR spectra of graphene oxide (GO), PEGylated graphene oxide (GO-PEG), and mouse liver tissue. Band of 3439 cm-1 (O-H stretching) is assigned to GO; 1437 cm-1 (-CH2 stretching) and 1291 cm-1 (C-O-C stretching) are ascribed to PEG polymer. Synchrotron FTIR images (b) and quantitative analysis (c) of the mouse kidney after i.v. injection of s-GOs and l-GOs at 24 h i.v. post-injection. ** p &lt; 0.001. Figure S7. Concentrations of s-GOs and l-GOs in stool samples after i.v. injection. Figure S8. Serum levels of the biomarkers of hepatic function after i.v. injection of 15 mg/kg s-GOs and l-GOs to mice (n = 5). Figure S9. Ultrastructure image of proximal tubule by TEM at 4 h after i.v. injection of lGOs. l-GOs deposited in peritubular capillary (a), tubular epithelial cytoplasm, (b) and tubular lumen (c). Clusters of GOs were indicated by red arrows. Figure S10. Cell viability assays of HK-2 cells following s-GO and l-GO treatments at 1, 10, 25, 50, 100, and 200 μg/mL doses for 24 h. ** p &lt; 0.01 and *** p &lt; 0.001 indicate the statistical difference between treated group and the controls. ### p &lt; 0.001 indicate the statistical difference between s-GO and l-GO treated group. Figure S11. The measurements of 24-hour ACR (albumin to creatinine ratio) in urine after s-GO and l-GO injection to mice at doses of 1, 5, 15 mg/kg bw. Figure S12. (a) Schematic of the detection of biomarkers and inflammatory cytokines in the kidneys for evaluation of kidney injury after GOs administration. (b) The mRNA expression level changes (i-vi) of kidney injury molecule-1 (Kim1) and neutrophil gelatinase-associated lipocalin (Ngal), and the inflammatory cytokines: Ccl2/Mcp1, Tnfα, IL1β and IL6 in kidney lysates at 48 h and day 7 after i.v. injection of 15 mg/kg bw s-GOs and l-GOs to mice (n = 3).