Figure 2 | Scientific Reports

Figure 2

From: Activation of mineralocorticoid receptor by ecdysone, an adaptogenic and anabolic ecdysteroid, promotes glomerular injury and proteinuria involving overactive GSK3β pathway signaling

Figure 2

Ecdysone induces glomerular molecular changes indicative of glomerulopathy. (A) Mice were treated as stated in Fig. 1. Kidney specimens were obtained on day 14 and prepared for Masson trichrome staining, immunohistochemistry staining for fibronectin and cleaved caspase-3 (Cleaved casp-3), and dual color fluorescent immunohistochemistry staining for synaptopodin (Synpo, green signal) and desmin (red signal). Bars = 20 µm. Ecdysone induced podocyte injury, marked by the reduced expression of synaptopodin and augmented expression of desmin in synaptopodin positive podocytes (yellow signal as indicated by the white arrowhead). Apoptotic cells, probed by positive staining for cleaved caspase-3, were predominantly located to the periphery of glomerular tufts that is consistent with podocyte localization (indicated by the black arrowhead). (B) Glomeruli were isolated from kidneys by the magnetic beads-based approach and homogenized for immunoblot analysis for indicated molecules, including fibronectin, α-smooth muscle actin (α-SMA), desmin, synaptopodin (Synpo), cleaved caspase-3 (Cleaved casp-3) and glyceraldehyde 3-phosphate dehydrogenase (GAPDH) (cropped blots are shown as indicated by the lines and full-length blots are included in the Supplementary Figure 1).

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