Fig. 5: Molecular features and function of regenerative Aundiff are dependent on growth factor signalling.
From: Distinctive molecular features of regenerative stem cells in the damaged male germline
a Upper panel: treatment regimen and analysis timeline for mice receiving multikinase inhibitor AD80 after BU. Lower panels: Representative wholemount IF of adult mice treated with BU then AD80 or vehicle daily for 5 days from D5 post-BU. Higher magnification grayscale images illustrating FOXO1 localisation in highlighted cells are shown. Graph shows FOXO1 distribution within E-CADHERIN + As/Apr at D10 (N, predominantly nuclear; N + C, nuclear and cytosolic; C, predominantly cytosolic) and P-value (χ2 test, two-sided) (n = 3 vehicle and n = 4 AD80 treated; >100 mm tubule length analysed per sample). b Representative wholemount IF of mice treated as in a. Higher magnification grayscale images of P-S6 in highlighted regions are shown. Graph indicates percentage of E-CADHERIN + As/Apr P-S6 + at D10 (n = 3 vehicle and n = 4 AD80). c, d Representative flow cytometry of testis from mice treated as in a at D10. Graphs indicate percentage of Aundiff (PLZF + c-KIT–) KI67 + and EOMES + in c and percentage of Aundiff (E-Cadherin+ c-KIT–) uPAR+ plus relative levels of EpCAM on Aundiff (median fluorescent intensity) in d (n = 4 mice per group). e Representative wholemount IF grayscale images illustrating SALL4 + populations in tubules from mice treated as in a at D30. Images taken along tubule length (n = 3 mice per group, >8 cm tubule length analysed per vehicle-treated control and >14 cm per AD80-treated animal). Insets show magnified regions of highlighted areas. Asterisks: tubule regions lacking SALL4 + germ cells and spermatogenesis. f Upper graph: Percentage of tubule length containing SALL4 + cells and spermatogenesis (n = 3 mice per group). Lower graph: Abundance of GFRα1+ spermatogonia per mm tubule length within spermatogenic regions (n = 4 mice per group). Dashed lines in a, b: tubule outline. Scale bars: 50 μm in a, b and 1 mm in e. Data presented as mean values ± SEM in b, c, d, f. Significance by two-tailed unpaired t test in b–d, f. Source data are provided as a Source Data file.
