Journal: iScience

Article Title: Caloric restriction protects from acute and chronic kidney injury by inhibiting monocyte recruitment

doi: 10.1016/j.isci.2025.113094

Figure Lengend Snippet: FMD reduces renal damage 14 and 35 days after AA injection (A and B) (A) Blood urea nitrogen (BUN) and (B) serum creatinine values at serial time points in BALB/c male mice injected with AA on ad lib diet ( n = 13) or FMD ( n = 18). (C) Representative brightfield images (H&E staining) of ad lib and FMD mice cortical tubular sections at 14 and 35 days after AA injection. A scale bar representing 100 μm is shown in the lower right corner. (D and E) Bar plots of histologic scores of tubular injury and inflammation severity at 14 days ( ad lib , n = 7; FMD, n = 8) and at 35 days ( ad lib , n = 11; FMD, n = 9) after AA injection. (F and G) Representative immunofluorescence (IF) images of KIM-1 (red), LTL (green), and DAPI (blue) cortical tubular staining and data quantification in ad lib and FMD mice at 14 days after AA injection ( ad lib , n = 8; FMD, n = 8). A scale bar representing 100 μm is shown in the lower right corner. (H and I) Representative IF images of KIM-1 (red), LTL (green), and DAPI (blue) cortical tubular staining and data quantification in ad lib and FMD mice at 35 days after AA injection ( ad lib , n = 8; FMD, n = 8). A scale bar representing 100 μm is shown in the lower right corner. Repeated measures ANOVA model was used to assess statistical significance at different time points (A and B). t test was used to compare distributions between groups at the same time point. Data are represented as mean ± SEM ○: p < 0.05 vs. baseline; ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, and ∗∗∗∗ p < 0.0001 vs. ad lib at the same time point; ns: not significant.

Article Snippet: Serum Blood Urea Nitrogen (BUN) was quantified in serial blood collections using colorimetric detection kit from Thermo Scientific according to the manufacturers’ instructions (Urea Nitrogen (BUN) Colorimetric Detection Kit, Product# EIABUN, Thermo Fisher Scientific).

Techniques: Injection, Staining, Immunofluorescence

Journal: iScience

Article Title: Caloric restriction protects from acute and chronic kidney injury by inhibiting monocyte recruitment

doi: 10.1016/j.isci.2025.113094

Figure Lengend Snippet: Renal damage, circulating monocytes, and renal infiltrating monocytes at 14 days after AA injection with or without CCR2i (A and B) (A) Blood urea nitrogen (BUN) and (B) serum creatinine values at serial time points after AA injection ( ad lib + vehicle n = 10, FMD + vehicle n = 7, ad lib + CCR2i n = 8, FMD + CCR2i n = 9); mice were injected with CCR2i or vehicle every day starting from the day of AA injection till day 14. (C) Representative brightfield images (H&E) of ad lib + vehicle, FMD + vehicle, ad lib + CCR2i and FMD + CCR2i of kidney cortical tubular sections at day 14 after AA injection. A scale bar representing 100 μm is shown in the lower right corner. (D) Bar plot of histologic scores of tubular injury and inflammation severity at 14 days ( ad lib + vehicle n = 7, FMD + vehicle n = 8, ad lib + CCR2i n = 4, FMD + CCR2i n = 5). (E) Circulating CD45 + CD11b + myeloid cell changes (%) over baseline at 7 and 14 days ( ad lib n = 10, FMD n = 7, ad lib + CCR2i n = 8, FMD + CCR2i n = 9). (F) Kidney quantification of CD45 + immune cells at 14 days ( ad lib n = 7, FMD n = 9, ad lib + CCR2i n = 8, FMD + CCR2i n = 9). (G) Kidney quantification of CD45 + CD11b + myeloid cells at 14 days ( ad lib n = 7, FMD n = 9, ad lib + CCR2i n = 8, FMD + CCR2i n = 9). (H) Kidney quantification of CD45 + CD11b + Ly6G − monocyte cells at 14 days ( ad lib n = 7, FMD n = 9, ad lib + CCR2i n = 8, FMD + CCR2i n = 9). (I) Kidney quantification of CD45 + CD11b + F480 + macrophage cells at 14 days ( ad lib n = 7, FMD n = 9, ad lib + CCR2i n = 8, FMD + CCR2i n = 9). (J) Kidney quantification of CD45 + CD11b + Ly6G − Ly6C high pro-inflammatory monocyte cells at 14 days ( ad lib n = 7, FMD n = 9, ad lib + CCR2i n = 8, FMD + CCR2i n = 9). (K) Kidney quantification of CD45 + CD11b + Ly6G − Ly6C low anti-inflammatory monocyte cells at 14 days ( ad lib n = 7, FMD n = 9, ad lib + CCR2i n = 8, FMD + CCR2i n = 9). (L) Kidney quantification of CD45 + CD11b + Ly6G + neutrophil cells at 14 days ( ad lib n = 7, FMD n = 9, ad lib + CCR2i n = 8, FMD + CCR2i n = 9). Repeated measures ANOVA model was used to assess statistical significance at different time points. One-way ANOVA was used to compare the 4 groups at the same time point. Data are represented as mean ± SEM ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, and ∗∗∗∗ p < 0.0001 vs. ad lib at the same time point; ns not significant.

Article Snippet: Serum Blood Urea Nitrogen (BUN) was quantified in serial blood collections using colorimetric detection kit from Thermo Scientific according to the manufacturers’ instructions (Urea Nitrogen (BUN) Colorimetric Detection Kit, Product# EIABUN, Thermo Fisher Scientific).

Techniques: Injection

Journal: iScience

Article Title: Caloric restriction protects from acute and chronic kidney injury by inhibiting monocyte recruitment

doi: 10.1016/j.isci.2025.113094

Figure Lengend Snippet: FMD-induced renal recovery after AKI in ad lib and FMD mice (A and B) (A) Blood urea nitrogen (BUN) and (B) serum creatinine values at serial time points after AA injection in ad lib ( n = 6) and FMD ( n = 8) BALB/c male mice until day 35. (C) Representative brightfield images of ad lib and FMD mice cortical tubular sections at different time points (upper quadrants H&E staining, lower quadrants Masson trichrome staining). A scale bar representing 100 μm is shown in the lower right corner. (D) Bar plot of histologic scores of tubular injury, inflammation, and fibrosis severity at 35 days ( ad lib , n = 6 and FMD, n = 8). (E, G, and I) Representative IF images of KIM-1 (orange), VCAM-1 (magenta), fibronectin (red) LTL (green) and DAPI (blue) cortical tubular staining in ad lib and FMD mice at 35 days. A scale bar representing 100 μm is shown in the lower right corner. (F, H, and J) KIM-1, VCAM-1, and fibronectin MFI quantification in cortical tubules (left) and % of cortical tubuli positive (right) (positivity assessed by comparing tubular KIM-1 MFI with unstained control) at 35 days ( ad lib , n = 8 and FMD, n = 8). Repeated measures ANOVA model was used to assess statistical significance at different time points (A and B). t test was used to compare distributions between groups at the same time point. Data are represented as mean ± SEM ○: p < 0.05 vs. baseline, ○○: p < 0.01 vs. baseline; ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, and ∗∗∗∗ p < 0.0001; ns, not significant.

Article Snippet: Serum Blood Urea Nitrogen (BUN) was quantified in serial blood collections using colorimetric detection kit from Thermo Scientific according to the manufacturers’ instructions (Urea Nitrogen (BUN) Colorimetric Detection Kit, Product# EIABUN, Thermo Fisher Scientific).

Techniques: Injection, Staining, Control

Journal: iScience

Article Title: Caloric restriction protects from acute and chronic kidney injury by inhibiting monocyte recruitment

doi: 10.1016/j.isci.2025.113094

Figure Lengend Snippet: Ad lib and FMD diet in B6 mice receiving AA and in BALB/c mice treated with folic acid (A and B) (A) Blood urea nitrogen (BUN) and (B) serum creatinine values at day 7 and day 14 after AA injection in ad lib ( n = 6) and FMD ( n = 6) B6 male mice. (C) Representative brightfield images of ad lib and FMD mice cortical tubular sections (H&E staining). A scale bar representing 100 μm is shown in the lower right corner. (D) Bar plot of histologic scores of tubular injury and inflammation at 14 days in ad lib ( n = 6) and FMD ( n = 6). (E and F) (E) Blood urea nitrogen (BUN) and (F) serum creatinine values at day 3 and day 7 after folic acid injection in ad lib ( n = 7) and FMD ( n = 6) BALB/c male mice. (G) Representative brightfield images of ad lib and FMD mice cortical tubular sections (H&E staining). A scale bar representing 100 μm is shown in the lower right corner. (H) Bar plot of histologic scores of tubular injury and inflammation at 7 days in ad lib ( n = 6) and FMD ( n = 6). Repeated measures ANOVA model was used to assess statistical significance at different time points (A, B, E, and F). t test was used to compare distributions between groups at the same time point. Data are represented as mean ± SEM ○: p < 0.05 vs. baseline, ○○: p < 0.01 vs. baseline; ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, and ∗∗∗∗ p < 0.0001; ns, not significant.

Article Snippet: Serum Blood Urea Nitrogen (BUN) was quantified in serial blood collections using colorimetric detection kit from Thermo Scientific according to the manufacturers’ instructions (Urea Nitrogen (BUN) Colorimetric Detection Kit, Product# EIABUN, Thermo Fisher Scientific).

Techniques: Injection, Staining