Table 1.
qPCR primers used in this study.
Fig 1.
Organ and body weight and serum biochemistries after HPD.
(A) 12-week old WT or Pin1 KO mice were fed HPD for 4, 8, 12 weeks before analysis. (B) Mouse body weights were measured at the beginning and end of normal diet (-) and HPD (+), and expressed as % increase compared to the weight at the beginning of diet given. * denotes p<0.01 between treatments with 6 mice/group. (C) and (D) Serum from each mouse was collected at the end of diet (12 weeks) and measured for the indicators of kidney function. (E) Kidney weight at the end of diet.
Fig 2.
Serum from each mouse were collected at the end of feeding (12 weeks) and measured for the electrolyte concentration (A) (B) (C) and the amount of Vitamin D (D). ** in (B) and (D) denotes p<0.05 between genotypes with (+) or without (-) HPD. * in (A) denotes p<0.05 between treatments (+ vs. -) in each genotype. All data were analyzed by student t-test with 6 mice each group.
Fig 3.
Collagen deposition by HPD is decreased by Pin1 deficiency.
(A) Kidney sections from 12-week HPD (+) mice were stained with Von Kossa for calcium-phosphate containing salts (dark) and the positive area in outer medulla was quantified as shown and described in the Methods. (B) and (C) Kidney sections from 8 and 12-week control diet (-) and HPD (+) mice were stained with Masson’s trichrome (left) for total collagen deposition. The intensity of blue color was quantified and shown in bar graphs (right). FS: free surface of kidney. ** denotes p<0.05 between genotypes by student's t-test with 6 mice each group.
Fig 4.
Renal Expression of ECM mRNAs.
(A) and (B) Kidney RNA from 8- and 12-weeks HPD mice was subjected to quantitative PCR. * denotes p<0.05 between genotypes with HPD (+) by student's t-test with 6 mice each group. (C) Whole kidney lysates from 12-weeks control and HPD mice were subjected to immunoblots with antibodies shown. The lysates were from 3–6 animals each group. Phosphorylated proteins were normalized to total proteins, and shown in bar graph (right). N.S.—not significant.
Fig 5.
HPD-induced kidney Inflammation is attenuated by the Pin1 deficiency.
(A) The expression of inflammasome component (NLRP3) and pro-inflammatory cytokines (IL-1β and TNF-α) in kidney from 12-weeks HPD (+) and control mice (-) were analyzed by quantitative PCR. (B) Macrophage infiltration was assessed by the expression of CD68. (C) Macrophages (left) were stained with anti-F4/80 and the density of brown color (arrow-heads) in the outer medulla was quantified (right) from 3 fields (20x) each slide. * denotes p<0.05 between genotypes with HPD with 6 mice per group.