Fig. 4: Spatial distribution of proliferating cells relative to necrotic sites.
From: Longitudinal tracking of acute kidney injury reveals injury propagation along the nephron
a, b Serial in vivo 2-photon microscopy images of representative Mid regions in CycB1-GFP reporter kidneys at indicated days after partial IRI. Note localized GFP expression (arrowheads) around necrotic sites (⊥) in PT-S1 segments while GFP expression arises also in remote distance of necrotic sites (⊥) in PT-S2. Scale bar: 50 μm. c Cumulative volumetric quantification of GFP-expressing nuclei (% of total nuclei/ segment) determined from days 1–3 after partial IRI for PT-S1, PT-S2, and DCT/CD, which arose either in proximity (<25 μm) or distant localization (>25 μm) from any propidium iodide (PI)+ cell detected on day 0 (n = 145, 184 and 37 segments from 4 mice for PT-S1, PT-S2, and DCT/CD, respectively); mean ± 95% CI with scatterplot. Statistical test: two-sided paired t-test with p < 0.05 considered significant (Supplementary Extended Statistics.k). d Same dataset as in (c) displayed to reveal distinct locations of proliferating tubule cells in different tubule segments over days after partial IRI (n = 4 partial IRI mice; detailed information on segment number/group provided in Supplementary Table 3); mean ± 95% CI with scatterplot. Statistical test: linear mixed-effect model, p-values from two-sided test (Supplementary Extended Statistics.m). e Volumetric quantification of tubular GFP expression (% of total nuclei/ segment) clustered by necrotic thresholds (n = 223, 140, 157 PT-S1 segments and 250, 237, 103 PT-S2 segments for non-necrotic, moderate, and severe, respectively, from 6 mice); mean ± 95% CI with scatterplots. Statistical test: linear mixed-effect model, p-values from two-sided test (Supplementary Extended Statistics.n). f Linear regression of PI+ nuclei (% of total nuclei/segment) at day 0 and cumulative GFP+ nuclei (% of total nuclei/ segment) from days 1, 2, and 3 for PT-S1 and PT-S2 segments (n = 144 and 180 from 4 mice). Fit (slope*x + intercept, plotted with 95% CI), R2 and Pearson’s coefficient (r) are reported for both. #: significant difference of slope and intercept from zero. *: significance test across groups, p-values from two-sided test. */#: p < 0.05; **/##: p < 0.01; ***/###: p < 0.001.
