Figure 4.

ANXA1sp treatment improves markers of mitochondrial integrity. Mice were treated with either Vehicle or ANXA1sp 1 h prior to ischemia, subjected to 33 min of unilateral ischemia and contralateral nephrectomy, and then re-injected with Vehicle or ANXA1sp 1 h after reperfusion. Kidney tissues were harvested at 24 h after reperfusion. (A) Protein levels of dynamin-related protein (Drp)1 were determined by Western blot with densitometry shown in the graph below. (B) Western blot for LC3 I/LC3 II showing protein levels of indicated proteins with densitometry shown in the graph below. (C) Western blot for Pink1 and Parkin with densitometry shown in (D) and (E), respectively. Graphs display mean +/-SEM of densitometry of protein normalized to tubulin. Statistical significance determined by two-way ANOVA with Sidak post-test (n = 3 samples for Sham groups, n = 6 samples for I/R groups; ^*p < 0.05, ^***p < 0.001). (F) Confocal immunofluorescence micrographs of the kidney cortex (left panels) and corticomedullary junction (right panels). Images show confocal micrographs of kidney sections stained for LC3 (red), citrate synthase (green), and nuclei are counterstained with DAPI. Arrows denote overlay of LC3 and citrate synthase (yellow/orange fluorescence). Note the increased citrate synthase expression and LC3 puncta in corticomedullary region of ANXA1sp-treated mice. Magnification = 600x. Data are representative of three sets of mice.