Fig 2: PAX8 expression in murine OSE cells increases migration and proliferation(A) Murine ovarian surface epithelium (MOSE) cells with forced PAX8 expression were significantly faster at healing a wound after 3 days (n = 3). (B) MOSE-PAX8 cells have increased migration through a Boyden chamber as depicted through the xCELLegence assay® (n = 3) in 20 hours. (C) No change in migration over a wound after 2 days in MOEsiPAX8 cells (n = 4). (D) Representative western blot demonstrating increased expression of N-cadherin and Fibronectin after PAX8 expression in MOSE cells. (E) MOSE-PAX8 cells have a more mesenchymal, elongated morphology when compared to MOSE-Neo. (F) MOSE-NEO and MOSE-PAX8 cells (1 × 107 cells/mouse) were intraperitoneally (IP) injected into nude mice, and internal gross anatomy was examined for tumors after 6 months. Data represent mean ± SD. Significance is represented by * for p = 0.05.

Fig 3: Morphological changes in PECs in diabetic db/db mouse. (a) Representative images of PAX8-LTA costaining in the 12-week-old db/m and db/db mice. The magnified view of the boxed region in the upper panel is shown in the lower panel. The dot plot graph shows the mean nuclear volume of PECs. (b) Representative images of PAS staining (upper) and LTA staining (lower) in the 12-week-old db/m and db/db mice. The dot plot graph shows the average of semiquantitative cuboidal PEC score in the db/m and db/db mice. (c) Immunostaining for CD44 in kidneys from the experimental mice groups (i.e., db/m, db/db, saline-treated, and adriamycin-treated). (d) Representative PECs (arrows) on transmission electron microscopy at 12 weeks of age in each mouse group. The dot plot graphs show the temporal changes in the cellular volume of PECs. (e,f) Temporal changes in the GBM thickness (e) and BBM thickness (f) in the db/m and db/db mice. (g) Representative PECs (arrows) on transmission electron microscopy at 24 weeks of age in the db/m and db/db mice. Arrowheads indicate the vacuole in PECs. The right inserts are the views on higher magnification. Quantitative analysis of the number of vacuoles per cell is shown in the lower panel. (h) A binucleated PEC is identified in db/db mice at 12 weeks of age but not in db/m mice. Arrows indicate PECs. PECs, parietal epithelial cells. *P < 0.05 vs. db/m mice. Scale bars: (a up) 20 µm, (a low) 5 µm, (b,c) 20 µm and (d,g,h) 5 µm.

Fig 4: Renal phenotypes in the three Cre Wt1 conditional models. (A-D) H&E-stained E18.5 embryonic kidneys. (A) Wt1co/co. (B) Nes-Cre Wt1co/co. (C) Wnt4+/CreGFP Wt1co/co. (D) Pax8+/Cre Wt1co/co. CM, cap mesenchyme; CB, comma-shaped body; SB, S-shaped body; UB, ureteric bud; PT, proximal tubule; DT, distal tubule; EM, expanded mesenchyme; IPT, immature proximal tubule. Scale bars: 50 µm. (E) Macroscopic view of Wilms’ tumour in single surviving Nes-Cre Wt1co/co mouse. Scale bar: 10 mm. (F) Same tumour as in E with left kidney from the same mouse. Scale bar: 10 mm. (G) H&E staining of the tumour in E and F. Scale bar: 500 µm.

Fig 5: Pax8 is not expressed in the developing mouse pancreas or in mature islets. Immunostaining of pancreatic consecutive sections using either the polyclonal (left panels) or monoclonal (right panels) Pax8 antibody, or the Pax6 antibody (middle panels) reveals an overlapping signal between Pax6 and the polyclonal Pax8 antibodies. The monoclonal Pax8 antibody does not stain mouse islets at any studied age. a–c E17.5, d–f Pup and g–i adult mice. Scale bar 50 µm