File S1 - Targeting JNK by a New Curcumin Analog to Inhibit NF-kB-Mediated Expression of Cell Adhesion Molecules Attenuates Renal Macrophage Infiltration and Injury in Diabetic Mice

A supplemental in vivo study to evaluate the renal-protective effects of C66 in STZ-induced diabetic mice. File S1 consists of Materials and Methods, and Supplemental Table and Figures (Table S1, Figure S1–S3). Table S1 in File S1. Primer sequences for real-time quantitative PCR. Figure S1 in File S1. C66 and SP600125 treatment attenuated the over-expression of adhesion molecules in diabetic mouse kidneys. A–C. The total RNA in kidney tissues of six groups in the supplemental in vivo study was extracted respectively, and was performed for RT-qPCR analysis to detect the mRNA level of VCAM-1 (A), ICAM-1 (B), and MCP-1 (C). The mRNA levels (mean ± SD) are expressed as a ratio of β-actin. n = 5–8/group). D–F. The total protein in kidney tissues of six groups in the supplemental in vivo study was extracted respectively, and was performed for Western blotting analysis to detect the protein level of VCAM-1 (D), ICAM-1 (E), and MCP-1 (F). The column figures show the normalized optical density from the blot data (4–6 mice in each group, * pFigure S2 in File S1. C66 and SP600126 treatment decreased renal injuries in diabetic mice in the supplemental in vivo study. A. Plasma creatinine levels were determined and normalized by that of one mouse in Con group (*pFigure S3 in File S1.. A schematic illustration for the prevention of C66 from HG-induced macrophage infiltration and renal injury. (DOC)