Extended Data Fig. 1: Renal H-scan accurately tracks the development of mouse kidney fibrosis.

(a) Schematic of mouse kidney imaging experiments. (b) Representative images of PSR-stained sections from sham kidneys as well as fibrotic kidneys on day 7 and day 14 following UUO surgery. Black scale bar: 100 µm. (c) Representative B-mode and (d) H-scan images of sham kidneys and kidneys at days 7 and 14 after UUO surgery. The H-scan maps are shown for the whole kidney and the outer (cortex) and inner regions (medulla). White scale bar: 2 mm. (e) H-scan-based fibrosis estimates (% red pixel density) of whole kidney, outer kidney, and inner kidney regions based on images of left kidneys of sham mice and day 7 and day 14 post-UUO mice (n = 5/group). For the whole kidney region of interest, a one-way ANOVA revealed a statistically significant difference between the groups F(2, 14) = 7.11, p = 0.0092, with a large effect size η = 0.54. Post-hoc analyses with Tukey’s Honestly Significant Difference analysis, with Bonferroni-corrected significance revealed a significance difference in H-scan %red between Sham vs. Day 7 (p = 0.034), Day 7 vs. Day 14 (p = 0.097), and Sham vs. Day 14 (p = 0.0073). The same analysis and reporting was performed for the outer and inner kidney regions of interest. For the outer kidney region of interest, the H-scan ANOVA statistical results are: F(2, 14) = 6.00, p = 0.016, η = 0.50, Sham vs. Day 7 p = 0.038, Day 7 vs. Day 14 p = 0.011, Sham vs. Day 14 p = 0.013. For the inner kidney region of interest, the H-scan ANOVA statistical results are: F(2, 14) = 3.81, p = 0.042, η = 0.39, Sham vs. Day 7 p = 0.039, Day 7 vs. Day 14 p = 0.037, Sham vs. Day 14 p = 0.042. Fibrotic burden measurements using gold standard (f) PSR, (g) type I collagen, and (h) α-SMA stains. For PSR staining, the ANOVA statistical results are: Whole kidney: F(2, 14) = 26.21, p = 4.18e-5, η = 0.81, Sham vs. Day 7 p = 0.0070, Day 7 vs. Day 14 p = 0.020, Sham vs. Day 14 p = 4.05e-5; Outer kidney: F(2, 14) = 21.24, p = 1.14e-, η = 0.78, Sham vs. Day 7 p = 0.0017, Day 7 vs. Day 14 p = 0.023, Sham vs. Day 14 p = 0.00010; Inner kidney: F(2, 14) = 15.71, p = 4.46e-4, η = 0.72, Sham vs. Day 7 p = 0.0066, Day 7 vs. Day 14 p = 0.026, Sham vs. Day 14 p = 0.0004. For type I collagen staining, the ANOVA statistical results are: Whole kidney: F(2, 14) = 10.92, p = 0.0020, η = 0.54, Sham vs. Day 7 p = 0.0085, Day 7 vs. Day 14 p = 0.0093, Sham vs. Day 14 p = 0.0014; Outer kidney: F(2, 14) = 9.95, p = 0.0028, η = 0.62, Sham vs. Day 7 p = 0.0061, Day 7 vs. Day 14 p = 0.019, Sham vs. Day 14 p = 0.0021; Inner kidney: F(2, 14) = 5.71, p = 0.018, η = 0.49, Sham vs. Day 7 p = 0.0068, Day 7 vs. Day 14 p = 0.0079, Sham vs. Day 14 p = 0.017. For α-SMA staining, the ANOVA statistical results are: Whole kidney: F(2, 14) = 28.10, p = 2.97e-5, η = 0.82, Sham vs. Day 7 p = 0.00050, Day 7 vs. Day 14 p = 0.016, Sham vs. Day 14 p = 2.81e-5; Outer kidney: F(2, 14) = 12.29, p = 0.0012, η = 0.67, Sham vs. Day 7 p = 0.018, Day 7 vs. Day 14 p = 0.0286, Sham vs. Day 14 p = 0.0010; Inner kidney: F(2, 14) = 122.99, p = 1.01e-8, η = 0.95, Sham vs. Day 7 p = 7.87e-8, Day 7 vs. Day 14 p = 0.021, Sham vs. Day 14 p = 1.72e-8. Correlation between H-scan based % red pixel density and histologic fibrosis scores derived from (i) PSR, (j) type I collagen, and (k) α-SMA staining for the whole kidney, outer and inner regions. Pearson correlation coefficients (r) and corresponding p values are provided for each comparison in each graph, all of which are statistically significant. Data are presented as mean +/− standard error. * p < 0.05. a created with BioRender.com.