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. 2012 Feb 13;61(3):728–738. doi: 10.2337/db11-0584

FIG. 5.

FIG. 5.

Catalase deficiency impaired peroxisomal biogenesis. A: At 4 and 10 weeks after the induction of diabetes, plasma FFAs (A) were measured as described in research design and methods. B–E: Renal PEX5 (B), PEX11-α (D), and Abcd3 (E) mRNA expressions were analyzed by real-time PCR. PEX5 protein (C) was measured by Western blot analysis. WT, nondiabetic control mice; WTDM, STZ-induced diabetic WT mice; CKODM, STZ-induced diabetic CKO mice. Data are mean ± SE of n = 7–8 mice per group. *P < 0.05 vs. WT; †P < 0.05 vs. WTDM at the same duration of diabetes; **P < 0.05 between two groups. F: The efficacy of siCat or hCAT in MMCs was analyzed by Western blot. MMCs were transfected with 20 nmol/L siCat or scramble siRNA for 24 h. Remaining siRNA were washed and then transfected with 100 ng hCAT. G–K: MMCs were then cultured under PhA±HG for 4 h (RNA) or 24 h (protein). NAC (5 mmol/L) was pretreated for 24 h where indicated. PEX5 mRNA (G and H) was measured by real-time PCR, and protein (J and K) was measured by Western blot. After 24 h, MMCs were fixed and then immunostained with anti-PEX5 (1:300) (I). Magnification ×630. siCat, MMCs transfected with 20 nmol/L siCat; hCAT, MMCs transfected with 100 ng pCMV-hCat; NAC (5 mmol/L); HG (30 mmol/L d-glucose); PhA (100 μmol/L). Data are mean ± SE of n = 4 experiments in the cell culture study. *P < 0.05 vs. scramble siRNA-transfected cells; †P < 0.05 vs. scramble siRNA-transfected cells cultured under PhA±HG; #P < 0.05 vs. siCat-transfected cells cultured under PhA+HG; n.s. not significant between two groups.