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. 2022 Jun 16;10:924751. doi: 10.3389/fcell.2022.924751

FIGURE 1.

FIGURE 1

ADR-Induced Proteinuria is Robust in the N and Modest in the J Substrain of C57BL/6 mice. (A) C57BL/6 J and N mice were injected with ADR (15 mg/kg/bw, i.v.) twice (Day 0 and Day 9). Urine, serum, and weight were collected throughout the study, and the mice were euthanized 6 weeks after the first injection, on Day 42, at which time the kidneys were harvested. (B) Representative urine gels with albumin bands are shown. Equal volumes of urine (5 µl) were analyzed by SDS-PAGE and Coomassie Blue staining. BSA (bovine serum albumin) standards were loaded at determined amounts and used to make a densitometry standard curve to determine albumin concentration. Urine from healthy un-injected mice were used as control. (C) Albumin/creatinine ratio was determined and plotted (*p < 0.05, **p < 0.01 vs. baseline of the same substrain, # p < 0.05, # p < 0.01 vs. J substrain at same week, Student t-test, n = 4/substrain). Dashed line parallel to X axis denotes the baseline albuminuria (D) Albumin/creatinine ratios plotted from urine samples from Week 0 and Week 6 for J and N substrains (*p < 0.05, **p < 0.01, Student t-test, n = 4/substrain). (E) Representative serum gels with albumin bands shown. Equal volumes of diluted serum (1 µl of 20X dilution) were analyzed by SDS-PAGE and Coomassie Blue staining. BSA (bovine serum albumin) standards were loaded at determined amounts and used to make a densitometry standard curve to determine albumin concentration. (F) Albumin values were determined and plotted (*p < 0.05 vs. baseline, Student one tailed t-test, n = 4/substrain). (G) Serum creatinine levels from Week 0 and Week 6 of both J and N substrains are plotted as measured by the enzymatic creatinine test kit (*p < 0.05). (H) Decrease in percentage body weight of both J and N substrains of mice at various time points compared to baseline, throughout the study are plotted. (*p < 0.05, **p < 0.01 vs. week 0 of same substrain, #p < 0.05 vs. J substrain of same week Student t-test, n = 4/substrain). Dashed line parallel to X axis denotes the baseline starting body weight.