Research Paper Volume 12, Issue 17 pp 17601—17624

Figure 6. A subset of newly generated podocytes from PEC origin (tdTomato⁺EGFP⁺) co-express nephrin in the glomerular tuft of aged mice. (A–D) Representative confocal images of tdTomato staining (red fluorescent protein, RFP) (red), EGFP⁺ (green) and nephrin staining (blue) in young and aged mice. The inserts show separate channels of the outlined glomeruli, with superscripts: 1=tdTomato, 2=EGFP, 3=nephrin and 4=merged. (A) Young mice (OC) showed that tdTomato⁺PECs (red) were observed along Bowman’s capsule (A¹) and genetically labeled podocytes (EGFP⁺ cells, green) (A²) co-express nephrin (blue) (A³), creating a cyan color (A⁴). (B) Aged mice (OC) showed that a subset of tdTomato⁺ PECs (labeled with red fluorescent anti-RFP) (B¹) co-expresses EGFP⁺ (green) (B²) and nephrin (blue) (B³) creating a pink/yellow color in glomerular tuft (marked with white arrow) (B⁴). (C) Young mice (JMC) showed that tdTomato⁺ staining PECs were observed along Bowman’s capsule (C¹) EGFP staining (C²) was detected in a typical podocyte distribution and overlaps with nephrin staining (C³), creating a cyan color (C⁴). (D) Aged mice (JMC) showed that tdTomato⁺ staining PECs (D¹) (marked with white arrows) have migrated onto the glomerular tuft and differentiated to a podocyte fate, co-staining with EGFP (green) (D²) and nephrin (blue) (D³) creating pink/yellow color (D⁴). Scale bars represent 25μm or 5μm (insets).