Chapter 5 88 to technical reasons, or from autopsy tissue. All tissue samples were obtained and handled in accordance with institutional guidelines and with the Code of Conduct regarding the responsible use of human tissues.(83) Patient characteristics are given in Table 3. Immunohistochemistry The mouse anti-dynamin antibody Hudy 1 (Upstate Biotechnology, catalog number 05-319, diluted 1:80) was used to detect dynamin. Hudy 1 is a monoclonal antibody that recognizes both dynamin 1 and dynamin 2. Hudy 1 recognizes the epitope of residues 822-838 within the proline-rich domain of dynamin. Anti-mouse Envision (Dako Cytomotion, Glostrup, Denmark) was used as a secondary antibody to detect the primary antibody. A rabbit anticathepsin L antibody (Abcam, ab203028, diluted 1:100) was used to detect cathepsin L. The results were analyzed by measuring the percent positive area in the glomeruli using the ImageJ digital image analysis program. No scarred glomeruli were included in these analyses. Statistical analyses Statistical analyses were performed using SPSS version 23.0 (IBM Corp., Armonk, NY). mRNA and protein levels were compared using one-way ANOVA (with Dunnett’s post hoc analysis when more than 3 groups were compared). The Student’s unpaired t-test was used when 2 or 3 groups were compared. Correlation analysis was performed using the Pearson correlation coefficient. Differences with a p-value < 0.05 were considered significant. Results Gene expression profiling between Dahl and SHR rats First, we investigated differences in gene expression between Dahl rats and SHR rats using microarray analysis. With respect to cytoskeleton-related genes, we found 27 genes that were differentially regulated between Dahl rats and SHR rats (Table 1). Of these 27 genes, 25 were significantly upregulated (including Dnm1), and two genes (Spna1 and Pmfbp1) were downregulated.
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