Figure 8. A subset of newly generated podocytes from PEC origin (tdTomato+EGFP+) de novo express VEGF-A in the glomerular tuft of aged mice. (A–D) Representative confocal images of tdTomato+ (red), EGFP+ (green) and VEGF-A (blue, podocyte specific marker) in young and aged mice. The inserts show separate channels of the outlined glomeruli, with superscripts: 1=tdTomato, 2=EGFP, 3=VEGF-A and 4=merged. (A) Young mice (OC) showed that tdTomato+PECs were observed along Bowman’s capsule (A1). EGFP reporter (A2) overlaps with VEGF-A staining (A3) and creates a cyan color (A4). (B) Aged mice (OC) showed that a subset of differentiated tdTomato+ PECs (red) (B1) co-expresses EGFP+ (green) (B2) and VEGF-A (blue) (B3) creating a yellow/white color in the glomerular tuft (white arrows) (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 VEGF-A 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 VEGF-A (blue) (D3) creating pink/yellow color (D4). Scale bars represent 25μm or 5μm (insets).