Fig. 2: CDKL5 activity increases in renal tubular epithelial cells during AKI.

a–c Bilateral renal ischemia was induced in male wild-type (C57BL/6) mice for 30 min followed by reperfusion for indicated timepoints. Blood urea nitrogen, serum creatinine, and histological analysis (H&E staining) were used to examine renal function and damage. d–f C57BL/6 mice were treated with cisplatin (30 mg/kg, intraperitoneal injection), and BUN, serum creatinine, and histological analysis were conducted at the indicated timepoints. g Representative H&E staining depicting tubular damage (indicated by asterisk) in both ischemic and cisplatin-treated mice. The graphs (a–f) represent data from a single experiment (n = 5 biologically independent samples), from one out of three independent experiments, all producing similar results. h Renal tissues from control, ischemic, and cisplatin-treated mice were used for western blot analysis of indicated proteins. Data presented are representative of five independent experiments, which yielded similar results. i–k Cdkl5 was immunoprecipitated from the kidneys of control, ischemic, and cisplatin-treated mice, followed by in vitro kinase assays. The representative western blots show the levels of Cdkl5 immunoprecipitated from tissue samples. The graphs represent data from a single experiment (n = 6 biologically independent samples), from one out of four independent experiments, all producing similar results. l Ggt1-Cre mice were crossed with ROSA^mT/mG mice to generate transgenic mice that express membrane-localized EGFP in renal tubular epithelial cells. A representative image shows EGFP expression in renal tubular cells. Arrows with dotted lines indicate tubular cells, while arrows with solid line show the glomerulus. m Schematic representation of the procedure used to isolate EGFP-positive renal epithelial cells. n Cdkl5 immunoprecipitation and in vitro kinase assay from indicated cells. The graph (n = 4) is representative of two independent experiments. In all the bar graphs, experimental values are presented as mean ± s.e.m. The height of error bar = 1 s.e. and p < 0.05 was indicated as statistically significant. One-way ANOVA followed by Dunnett’s (a–f and i, j) or Tukey’s multiple-comparison test (n) was carried out, and statistical significance is indicated by *p < 0.05, **p < 0.01, ***p < 0.001. Scale bar (g and i): 100 µm. Source data are provided as a Source Data file.