Figure 6: Refinement of reprogramming factors and conditions enables the generation of iNPlike cells from primary human proximal tubule cells (hRPTECs).
(A) Brightfield images of parental (untransfected) hRPTECs and hRPTEC iNPs that have undergone transfection with pT-SES, pT-TetON and pEF1-HA-m7pB followed by reprogramming. (B) qRT-PCR analysis of NP gene expression in hRPTEC iNP-like cells relative to parental hRPTECs. Data are presented as mean ± SEM. Statistical significance was determined by Student’s t-test (**P≤0.01, n = 3 biological replicates). (C) Immunofluorescence of the mouse kidney organoid assay demonstrating the contribution of hRPTEC iNP-like cells to developing nephrons marked by E-Cadherin (ECad; 692 green). Arrows depict examples of integrated hRPTEC iNP-like cells marked by human nuclear 693 antigen (HuNu; red) and DAPI (nuclei; blue). Scale bar represents 30 μm.