Journal: bioRxiv

Article Title: ZEB2 signaling is essential for ureteral smooth muscle cell differentiation and maintenance

doi: 10.1101/2025.02.23.639741

Figure Lengend Snippet: (A) ZEB2 (green) expression is absent in Zeb2 flox/flox ; Tbx18Cre + knockout mice ( Zeb2 cKO) as compared to wild-type littermate. (B) 3-week-old Zeb2 cKO mice were runted with smaller body sizes as compared to wild-type control littermates. (C) 3-week-old Zeb2 cKO mice developed hydroureter and hydronephrosis. (D) PAS staining of the kidney of a 3-week-old Zeb2 cKO mouse showing dysplastic kidney with dilated ureter and hydronephrosis (*). (E) Newborn (P0) Zeb2 cKO mice develop hydroureter and hydronephrosis. (F) Histology of a newborn (P0) Zeb2 cKO mouse ureter with hydroureter as compared to wild-type littermate controls. (G) Immunofluorescence staining of ACTA2 (smooth muscle cell marker) and UPKIII (urothelium marker) shows abnormal ureter development with significant ureter dilation in the newborn (P0) Zeb2 cKO mouse as compared to the wild-type littermate controls. u: ureter, k: kidney, UB: urinary bladder. Dotted lines encircle the ureter. Scale bars: 25 µm (A and G); 100 µm (D and F).

Article Snippet: Following primary antibodies were used: ZEB2 (1:100, HPA003456; Sigma-Aldrich, USA or SC-271984, Santa Cruz Biotechnology, USA), TBX18 (1:100, ab115262, Abcam, USA), E-cadherin (1:50, Clone 36/610181; BD Biosciences, USA), ACTA2 (1:200, A2547; Sigma-Aldrich, USA), Cytokeratin (1:100, C2562, Sigma-Aldrich, USA), TAGLN (1:200, ab14106, Abcam, USA), FOXD1 (1:100 dilution, sc-47585; Santa Cruz Biotechnology USA), POSTN (1:100, ab14041, Abcam, USA), SOX9 (1:200, AB5535, Millipore, USA), GFP (GFP-1020, 1:100, Aves Labs, USA or A11122, 1:250; Invitrogen, Carlsbad, CA).

Techniques: Expressing, Knock-Out, Control, Staining, Immunofluorescence, Marker

Journal: bioRxiv

Article Title: ZEB2 signaling is essential for ureteral smooth muscle cell differentiation and maintenance

doi: 10.1101/2025.02.23.639741

Figure Lengend Snippet: (A, B) Ureteral smooth muscle cells are formed in the wild-type mice at E15.5 specified by smooth muscle markers TAGLN (A) and ACTA2 (B). There are no TAGLN + and ACTA2 + ureteral smooth muscle cells in Zeb2 cKO mice. Pan-cytokeratin (Pan-cyto) is used to mark urothelial cells. (C, D) Compared to wild-type littermate controls, ureteral tunica adventitia cell layers labeled by POSTN+ and FOXD1+ markers are expanded in Zeb2 cKO mice. Scale bars: 10 µm (A-D).

Article Snippet: Following primary antibodies were used: ZEB2 (1:100, HPA003456; Sigma-Aldrich, USA or SC-271984, Santa Cruz Biotechnology, USA), TBX18 (1:100, ab115262, Abcam, USA), E-cadherin (1:50, Clone 36/610181; BD Biosciences, USA), ACTA2 (1:200, A2547; Sigma-Aldrich, USA), Cytokeratin (1:100, C2562, Sigma-Aldrich, USA), TAGLN (1:200, ab14106, Abcam, USA), FOXD1 (1:100 dilution, sc-47585; Santa Cruz Biotechnology USA), POSTN (1:100, ab14041, Abcam, USA), SOX9 (1:200, AB5535, Millipore, USA), GFP (GFP-1020, 1:100, Aves Labs, USA or A11122, 1:250; Invitrogen, Carlsbad, CA).

Techniques: Labeling