Figure 6. CA in adult mice sensitizes... | Rockefeller University Press

Figure 6.

$Figure 6. CA in adult mice sensitizes kidneys to renal injury. (A) H&E staining of control and Ksp-Plk4 kidneys isolated at P7 and P30. No prominent cysts were evident at these stages. (B) Kidney samples from P30 Ksp-Plk4 mice immunostained with anti-mCherry to identify cells expressing mChPlk4. There was robust expression of Plk4 in DBA-positive collecting ducts and CLC-K–positive distal tubules, but absent in LTL-positive proximal tubules, consistent with Ksp-Cre–mediated expression. Nuclei were stained with DAPI. (C) Immunostaining of P30 control and Ksp-Plk4 kidneys to mark centrosomes (γ-tubulin), cilia (acetylated α-tubulin), collecting ducts (DBA), and DNA (DAPI). In contrast to the normal centrosome number in control kidneys, CA is evident in Ksp-Plk4. White box denotes regions shown in insets. Arrowheads point to cells with multiple centrosomes and cilia. Bars (insets), 2 µm. (D) Quantification of CA in P30 WT and Ksp-Plk4 mice. n = 747 cells (WT) and 562 cells (Ksp-Plk4). (E) Quantification of the percentage of cells expressing zero, one, or more than one primary cilium in P30 Ksp-Plk4 mice containing excess centrosomes compared with cells with normal centrosome number. CA caused an increase in the fraction of superciliated cells. n = 274 cells (WT) and 172 cells (Ksp-Plk4). Statistical significance was determined using a two-tailed unpaired t test (*, P < 0.05). (F) H&E staining of kidneys after unilateral renal IRI. Ischemic injury was induced in 3-mo-old Ksp-Plk4 mice and littermate controls by clamping the left renal pedicle for 45 min. Kidneys were isolated 30 d after IRI. Injured kidneys of Ksp-Plk4 mice rapidly developed cysts, which were absent in the contralateral (uninjured) kidney, as well as the injured kidney of control mice. n = two mice (WT control) and five mice (Ksp-Plk4).$

CA in adult mice sensitizes kidneys to renal injury. (A) H&E staining of control and Ksp-Plk4 kidneys isolated at P7 and P30. No prominent cysts were evident at these stages. (B) Kidney samples from P30 Ksp-Plk4 mice immunostained with anti-mCherry to identify cells expressing mChPlk4. There was robust expression of Plk4 in DBA-positive collecting ducts and CLC-K–positive distal tubules, but absent in LTL-positive proximal tubules, consistent with Ksp-Cre–mediated expression. Nuclei were stained with DAPI. (C) Immunostaining of P30 control and Ksp-Plk4 kidneys to mark centrosomes (γ-tubulin), cilia (acetylated α-tubulin), collecting ducts (DBA), and DNA (DAPI). In contrast to the normal centrosome number in control kidneys, CA is evident in Ksp-Plk4. White box denotes regions shown in insets. Arrowheads point to cells with multiple centrosomes and cilia. Bars (insets), 2 µm. (D) Quantification of CA in P30 WT and Ksp-Plk4 mice. n = 747 cells (WT) and 562 cells (Ksp-Plk4). (E) Quantification of the percentage of cells expressing zero, one, or more than one primary cilium in P30 Ksp-Plk4 mice containing excess centrosomes compared with cells with normal centrosome number. CA caused an increase in the fraction of superciliated cells. n = 274 cells (WT) and 172 cells (Ksp-Plk4). Statistical significance was determined using a two-tailed unpaired t test (*, P < 0.05). (F) H&E staining of kidneys after unilateral renal IRI. Ischemic injury was induced in 3-mo-old Ksp-Plk4 mice and littermate controls by clamping the left renal pedicle for 45 min. Kidneys were isolated 30 d after IRI. Injured kidneys of Ksp-Plk4 mice rapidly developed cysts, which were absent in the contralateral (uninjured) kidney, as well as the injured kidney of control mice. n = two mice (WT control) and five mice (Ksp-Plk4).