Fig. 7

Pax2 lineage-positive cells proliferate, while other TEC endocycle and are persistently lost after AKI. a Schematic procedure. b–j FACS analysis shows mCherry+ and mVenus+ cells in total renal cells of Pax8/FUCCI2aR (b, d) and Pax2/FUCCI2aR mice (c, d) at T0, of Pax8/FUCCI2aR (e, g) and Pax2/FUCCI2aR mice (f, g) at IRI T2 and of Pax8/FUCCI2aR (h, j) and Pax2/FUCCI2aR mice (i, j) at IRI T30. A representative experiment out of 4 is shown. k Number of total FUCCI2aR cells in Pax8/FUCCI2aR and Pax2/FUCCI2aR mice at T0, IRI T2, IRI T30 (n = 4 in each group). Mann–Whitney test *p < 0.05. l Number of new FUCCI2aR cells in Pax8/FUCCI2aR and Pax2/FUCCI2aR mice at IRI T30 in comparison to IRI T2 (n = 4 in each group). Mann–Whitney test NS. m Percentage of Pax2/FUCCI2aR over Pax8/FUCCI2aR total cells at T0, IRI T2, IRI T30 (n = 4 in each group). Mann–Whitney test *p < 0.05. n Number of total mVenus+ cells mCherry+mVenus+ cells in Pax8/FUCCI2aR and Pax2/FUCCI2aR mice at T0, IRI T2, IRI T30 (n = 4 in each group). Mann–Whitney test NS. o, p Cell cycle distribution of mCherry+, mVenus+, and mCherry+mVenus+ cells in Pax8/FUCCI2aR (o) and Pax2/FUCCI2aR mice (p) at IRI T30. A representative experiment out of 4 is shown. q Percentage of endocycling cells in Pax2/FUCCI2aR (black column) and Pax8/FUCCI2aR mice (light gray column) at IRI T30 (n = 4 in each group). Mann–Whitney test *p < 0.05. r mCherry+ cells (red) and p-H3+ cells (blue) in LTA+ tubules in the cortex (white) of Pax8/FUCCI2aR mice at IRI T30 (n = 5). Arrows show mCherry+p-H3+ cells. r′ Detail of r. s mCherry+ cells (red) and mVenus+ cells (green) in LTA+ tubules in the cortex (white) of Pax8/FUCCI2aR mice at IRI T30 (n = 5). Arrow shows mVenus+ cell. t Percentage of endocycling cells (mCherry+p-H3+ cells) and cycling cells (mVenus+ cells) in the cortex of Pax8/FUCCI2aR mice at IRI T30 (n = 5). Mann–Whitney test *p < 0.05. u Percentage of endocycling (mCherry+p-H3+ cells) and cycling (mVenus+ cells) AQP2− cells in the cortex and in OSOM of Pax8/FUCCI2aR mice at IRI T30 (n = 5). Mann–Whitney test *p < 0.05. v Measurement of the cell surface area of mCherry+p-H3+ cells and of mCherry+p-H3− cells after staining with Phalloidin (white) with Image J software in Pax8/FUCCI2aR mice at IRI T30 (n = 4). w Cell surface area of mCherry+p-H3+ cells and of mCherry+p-H3− cells in Pax8/FUCCI2aR mice at IRI T30. n = at least 20 cells for each mouse (n = 4), Mann–Whitney test ***p < 0.001. Data are mean ± SEM. Scale bars 20 µm. T0 = day 0, IRI T2 = day 2 after IRI, IRI T30 = day 30 after IRI