Figure 3. CA results in decreased cell... | Rockefeller University Press

Figure 3.

$Figure 3. CA results in decreased cell proliferation, increased apoptosis, and reduced nephron endowment. (A) E15.5 and E17.5 kidney sections immunostained with anti-pHH3 to mark proliferating mitotic cells. Nuclei were stained with DAPI. (B) Quantification of pHH3-positive mitotic cells per unit area (mm2) in kidney sections from Hoxb7-Plk4, Six2-Plk4, and control embryos at E15.5, E17.5, and P0. There was a significant decrease in pHH3 foci/mm2 predominantly at E15.5, with no statistically significant differences at later time points. A minimum of four to five sections was scored per genotype at each developmental stage. A two-tailed unpaired t test was used for statistical analysis (*, P < 0.05). (C) Representative images from E17.5 Hoxb7-Plk4 and control mice stained for apoptotic cells (TUNEL), E-cadherin (UB tubules), and Six2 (MM). Nuclei were stained with DAPI. Apoptotic cells (white arrowheads) were evident in both UB and MM cells. (D) Graph illustrating the fraction of TUNEL-positive cells per unit area (mm2) in kidneys between E15.5 and E17.5. n = 5 kidneys per genotype for each time point. Statistical significance was determined using a two-tailed unpaired t test (*, P < 0.05). (E) Brightfield image of whole kidneys (top) and H&E staining of midsagittal sections (middle) from Hoxb7-Plk4, Six2-Plk4, and control littermates at P0, highlighting the small kidney phenotype at birth. Immunofluorescence images (bottom) of sections stained for Podocin to identify glomeruli. Nuclei were stained with DAPI. (F) Quantification of kidney–to–body weight ratio, highlighting a significant reduction in kidney size. n = 33 (WT), 7 (Hoxb7-Plk4), and 5 (Six2-Plk4). (G and H) Quantification of the mean number of glomeruli per section from H&E- (G) and Podocin-stained (H) sections. There was a significant decrease in the number of glomeruli in Hoxb7-Plk4 and Six2-Plk4 mice compared with control, indicative of reduced nephron endowment. n = 7 (WT) and 5 (Hoxb7-Plk4 and Six2-Plk4). Two-tailed unpaired t test was used for statistical analysis (*, P < 0.05).$

CA results in decreased cell proliferation, increased apoptosis, and reduced nephron endowment. (A) E15.5 and E17.5 kidney sections immunostained with anti-pHH3 to mark proliferating mitotic cells. Nuclei were stained with DAPI. (B) Quantification of pHH3-positive mitotic cells per unit area (mm²) in kidney sections from Hoxb7-Plk4, Six2-Plk4, and control embryos at E15.5, E17.5, and P0. There was a significant decrease in pHH3 foci/mm² predominantly at E15.5, with no statistically significant differences at later time points. A minimum of four to five sections was scored per genotype at each developmental stage. A two-tailed unpaired t test was used for statistical analysis (*, P < 0.05). (C) Representative images from E17.5 Hoxb7-Plk4 and control mice stained for apoptotic cells (TUNEL), E-cadherin (UB tubules), and Six2 (MM). Nuclei were stained with DAPI. Apoptotic cells (white arrowheads) were evident in both UB and MM cells. (D) Graph illustrating the fraction of TUNEL-positive cells per unit area (mm²) in kidneys between E15.5 and E17.5. n = 5 kidneys per genotype for each time point. Statistical significance was determined using a two-tailed unpaired t test (*, P < 0.05). (E) Brightfield image of whole kidneys (top) and H&E staining of midsagittal sections (middle) from Hoxb7-Plk4, Six2-Plk4, and control littermates at P0, highlighting the small kidney phenotype at birth. Immunofluorescence images (bottom) of sections stained for Podocin to identify glomeruli. Nuclei were stained with DAPI. (F) Quantification of kidney–to–body weight ratio, highlighting a significant reduction in kidney size. n = 33 (WT), 7 (Hoxb7-Plk4), and 5 (Six2-Plk4). (G and H) Quantification of the mean number of glomeruli per section from H&E- (G) and Podocin-stained (H) sections. There was a significant decrease in the number of glomeruli in Hoxb7-Plk4 and Six2-Plk4 mice compared with control, indicative of reduced nephron endowment. n = 7 (WT) and 5 (Hoxb7-Plk4 and Six2-Plk4). Two-tailed unpaired t test was used for statistical analysis (*, P < 0.05).