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. 2019 Jul 10;30(9):1659–1673. doi: 10.1681/ASN.2018121244

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Copyright © 2019 by the American Society of Nephrology

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Figure 5. — Castration led to decreases in renal EGFR expression and injury in male Dsk5 mice. (A) Male Dsk5 mice were castrated at 21 days of age and euthanized at 6 months of age. (B) Castration led to decreases in renal EGFR activation as indicated by phospho-EGFR (Y845) immunofluorescence staining, total renal EGFR expression, and p-ERK levels, a downstream signaling of EGFR activation. Original magnification, ×400 (scale bar, 20 µm). (C) Glomerular sclerosis, tubular dilation, and protein casts were apparent in sham male Dsk5 mice, whereas only mesangial expansion was seen in castrated male Dsk5 mice. Original magnification, ×160 (scale bar, 62.5 µm). (D) Loss of WT1 staining, a marker of podocytes, seen in male Dsk5 mice was significantly attenuated but was not completely prevented by castration (**P<0.01; ***P<0.001 versus wild-type group; ^††P<0.01 versus sham Dsk5 group; n=6 in each group). Original magnification, ×400 (scale bar, 25 µm). (E) Tubular injury seen in male Dsk5 mice was dramatically attenuated by castration as indicated by significant reduction of renal expression levels of KIM-1 and NGAL, two markers of tubular injury (***P<0.001 versus wild-type group; ^†††P<0.001 versus sham Dsk5 group; n=8 in wild-type group and n=7 in other groups). (F) Castration prevented progressive albuminuria seen in male Dsk5 mice (*P<0.05; **P<0.01; ***P<0.001 versus sham Dsk5 mice; n=8 in wild-type group and n=7 in other groups). (G) Castration significantly reduced tubulointerstitial fibrosis in male Dsk5 mice as indicated by Sirius Red staining (***P<0.001 versus wild-type group; ^†††P<0.001 versus sham Dsk5 group; n=6). Original magnification, ×160 (scale bar, 80 µm). (H) Castration decreased renal expression levels of α-SMA and CTGF in male Dsk5 mice. (I) Male Dsk5 mice had significant increases in renal expression levels of HB-EGF and amphiregulin (AREG), two ligands for EGFR, which were significantly attenuated by castration (**P<0.01, ***P<0.001 versus wild-type group; ^††P<0.01, ^†††P<0.001 versus sham Dsk5 group; n=8 in wild-type group and n=7 in other groups).

Figure 5. — Castration led to decreases in renal EGFR expression and injury in male Dsk5 mice. (A) Male Dsk5 mice were castrated at 21 days of age and euthanized at 6 months of age. (B) Castration led to decreases in renal EGFR activation as indicated by phospho-EGFR (Y845) immunofluorescence staining, total renal EGFR expression, and p-ERK levels, a downstream signaling of EGFR activation. Original magnification, ×400 (scale bar, 20 µm). (C) Glomerular sclerosis, tubular dilation, and protein casts were apparent in sham male Dsk5 mice, whereas only mesangial expansion was seen in castrated male Dsk5 mice. Original magnification, ×160 (scale bar, 62.5 µm). (D) Loss of WT1 staining, a marker of podocytes, seen in male Dsk5 mice was significantly attenuated but was not completely prevented by castration (**P<0.01; ***P<0.001 versus wild-type group; ^††P<0.01 versus sham Dsk5 group; n=6 in each group). Original magnification, ×400 (scale bar, 25 µm). (E) Tubular injury seen in male Dsk5 mice was dramatically attenuated by castration as indicated by significant reduction of renal expression levels of KIM-1 and NGAL, two markers of tubular injury (***P<0.001 versus wild-type group; ^†††P<0.001 versus sham Dsk5 group; n=8 in wild-type group and n=7 in other groups). (F) Castration prevented progressive albuminuria seen in male Dsk5 mice (*P<0.05; **P<0.01; ***P<0.001 versus sham Dsk5 mice; n=8 in wild-type group and n=7 in other groups). (G) Castration significantly reduced tubulointerstitial fibrosis in male Dsk5 mice as indicated by Sirius Red staining (***P<0.001 versus wild-type group; ^†††P<0.001 versus sham Dsk5 group; n=6). Original magnification, ×160 (scale bar, 80 µm). (H) Castration decreased renal expression levels of α-SMA and CTGF in male Dsk5 mice. (I) Male Dsk5 mice had significant increases in renal expression levels of HB-EGF and amphiregulin (AREG), two ligands for EGFR, which were significantly attenuated by castration (**P<0.01, ***P<0.001 versus wild-type group; ^††P<0.01, ^†††P<0.001 versus sham Dsk5 group; n=8 in wild-type group and n=7 in other groups).