Fig 2: Characterization of human podocytes derived from iPSCs in 12 days. A Immunostaining of iPSC-podocytes showing podocyte lineage markers PODXL (green), P-CADHERIN (green), MAFB (red), SYNPO (green), and F-actin (red) for cytoskeleton. Scale bar is 100 µm. B Pseudo-colored scanning electron microscopy image of an iPSC-podocyte in blue showing large cell bodies with foot-like projections and extracellular vesicles. Scale bar is 30 µm. C Transmission electron microscopy image of podocytes showing tight junction-like structures between cells (black arrows). Scale bar is 400 nm. D Flow cytometry histogram plots of the podocyte markers SYNPO and WT1 stained with a FITC and Texas Red secondary antibody (n = 3). E Quantitative analysis of podocyte markers such as PODXL, WT1, SYNPO, and NEPHRIN throughout the differentiation process was analyzed using (i) RT-PCR and (ii) Western blot quantification with ß-actin as control. *Indicates p < 0.05

Fig 3: Comparison of the protocol with existing protocols. A Schematic of the podocyte differentiation protocols compared including key media components. B(i) Comparison of the podocyte markers MAFB and PODXL expression using flow cytometry analysis (n = 3). (ii) Quantification of (Bi). C Immunostaining of iPSC-podocytes with markers PODXL, NPHS1 in the compared protocols (n = 3). Scale bar 100 µm. D Western blot analysis of the podocyte markers PODXL, SYNPO, NPHS1, and WT1 as well as loading control ß-actin in the compared protocols (n = 3)