Figure 5.

Tmbim6-knockout increases CsA-induced kidney injury and ER stress. (A) The mRNA expression of Tmbim6 was examined in whole kidney, renal cortex, and outer and inner medulla. Real-time PCR results from different sections of the kidney indicated that endogenous Tmbim6 mRNA levels were higher in the cortex region than in the outer and inner medullar regions. (B) In situ hybridization (ISH) analysis of Tmbim6, WT mice revealed clusters of brown punctate signals in most of the cells in the cortex and medulla, and signals were strongest in distal and proximal convoluted tubules. ((C)and D) WT mice and tmbim6^−/− mice were treated with 15 mg/kg CsA by intraperitoneal injection daily for 30 d, and blood samples were collected for measurements of serum urea and serum creatinine. Each value represents the mean ± SEM of 3 independent experiments. ^#, P < 0.05 CsA tmbim6^−/− mice vs. CsA WT mice. (E) Kidney tissue sections were used for PAS staining. Representative images are shown at × 600 magnification. (F) Immunoblotting of kidney lysates from WT mice and tmbim6^−/− mice treated with or without CsA was performed using anti-HSPA5, -DDIT3, and -ACTB antibodies. Densitometric analysis results are shown in the right panel. ^#, P < 0.05 CsA tmbim6^−/− mice vs. CsA WT mice. (G) Sirius Red staining was performed in the kidney tissue section to see the fibrosis.