Data for: Tcf21 as a Founder Transcription Factor in Specifying Foxd1 Cells to the Juxtaglomerular Cell Lineage

Supplemental Figures 1-5: 

Suppl Fig. 1: Tissue clearing and immunostaining of kidneys from Ren1dCre/+;Tcf21f/f and controls. Babb clearing, immunostaining with the pan-endothelial marker PECAM (CD31; red), and 3D imaging show no difference in vascular morphology between Ren1dCre/+;Tcf21f/f and control kidneys at E16.5. n=4, females. 

Suppl Fig. 2: Histology from 2-month-old Ren1dCre/+;Tcf21f/f and control kidneys. H&E (top) and Trichome (bottom) stainings reveal no gross phenotypic changes in Ren1dCre/+;Tcf21f/f kidneys compared with controls. Vascular wall and glomeruli morphology appear intact. n=16, females. Scale bar 100 μm. 

Suppl Fig. 3: Normal renin mRNA expression in Ren1dCre/+;Tcf21f/f kidneys at 2 months. In-situ hybridization (RNAscope). A-B. Renin expression (blue) is detected in the arteriolar walls (yellow arrows) and in the glomerulus hilum (black arrows) similarly in 2-month-old Ren1dCre/+; Tcf21f/f and control kidneys. C. The distribution of renin-expressing cells is similar in Ren1dCre/+; Tcf21f/f and controls. n=4, females. Scale bar 75μm (A), 100μm (B,C). 

Suppl Fig. 4: In-situ hybridization of Tcf21 and renin in E14.5 Ren1d+/+;Tcf21f/f kidneys. In-situ hybridization (RNAscope) shows no co-localization of Tcf21 (purple) and renin (blue) mRNA in E14.5 control kidneys. This time point was chosen because renin-expressing cells emerge around E13.5. n=2, sex not determined.

Suppl Fig. 5: Juxtaglomerular (JG) Cell Psuedotime Trajectory via scRNA-seq and scATAC-seq. A. UMAP visualization showing clustering of the entire GFP+ cell population isolated from Foxd1Cre/+;Rosa26mTmG/+ control mouse kidneys at E12, E18, P5, and P30 (n=6,191 cells). A total of 19 populations were identified. Populations representing stromal derivatives (clusters 2-16) are highlighted in red. B. Bubble plot demonstrating expression of key kidney cell lineage markers across the 19 clusters.  C. Cluster heatmap showing marker expression across all GFP+ clusters. D. UMAP visualization of the pseudotime trajectory mapping the differentiation form metanephric mesenchyme progenitors to mature renin-expressing JG cells (n=2,054 cells), shown by embryonic stage. Early populations with high Foxd1 expression serve as the starting point, and late populations with high Ren1/Akr1b7 expression were identified as endpoints. E-G. Feature plots visualizing Foxd1, Renin, and Akr1b7 expression levels across the entire GFP+ cell populations.

Supplemental Table 1: 

Suppl Table 1: Primer information (genotyping).