Figure 7. Effect of CCAAT enhancer binding protein beta (C/EBP-β) on ischemia-reperfusion (I/R)-induced renal dysfunction and apoptosis in mice.
(A,B) Lentivirus-mediated of C/EBP-β and pri-miR-16 in the kidney. The recombinant lentivirus carrying C/EBPβ or GFP as controls (1 × 109 viral particles per animal) was perfused into the wild-type kidney through renal artery. Two weeks after transduction, both C/EBP-β and pri-miR-16 mRNA level were assayed by real time polymerase chain reaction. (C) C/EBP-β is required for enhancing the I/R-induced renal dysfunction. Control (GFP or empty virus) or C/EBP-β mice underwent sham operation or I/R injury for 8 h. Blood urea nitrogen and creatinine levels, indicators for renal function, were measured. Data are means ± SEM (n = 5 in each group). *P < 0.05 and **P < 0.01, GFP versus C/EBP-β knockdown. (D) Apoptotic kidney cells in mice infused with or without C/EBP-β using in vivo TUNEL staining or kidney lysates were probed with specific antibody against the cleaved, active form of caspase-3. The full-length gels are presented in Supplementary Figure 7D. Colocalization of blue and brown staining in nuclei reflects apoptotic cells which are indicated with arrows. Bar = 50 μm. (E) Urinary miR-16 levels in mice with or without overexpression of C/EBP-β in the presence or absence of renal I/R.