Fig 1: Distinct NPHS2 variants resulted in differential trafficking abnormalities. (A)–(C) Kidney organoids immunostained for PODOCIN and endoplasmic reticulum (A, KDEL), Golgi (B, GM130), or late endosome (C, RAB7) markers showing partial colocalization in various subcellular structures for all variant cell lines at D7 + 17 (scale bar, 5 μm). DAPI stains nuclei. Images are adjusted to maximum value. (D) Relative quantification of PODOCIN subcellular compartment localization in control and variant glomeruli presented as the average modified Manders’ colocalization coefficient across different subcellular markers investigated per line. Values of the same subcellular marker should be compared across lines and not different subcellular markers within the same line.
Fig 2: Genetically engineered NPHS2 allelic series differentiated into kidney organoids. (A) Schematic of the PODOCIN protein showing the five missense variants characterized in this study, G92C, P118L, R138Q, R168H, and R291W. (B) Brightfield representative examples of kidney organoids differentiated from control and gene-edited cell lines at D7 + 17 (scale bar, 1000 μm). (C) Kidney organoids immunostained for glomeruli (NEPHRIN), proximal (Lotus tetragonolobus lectin, LTL), distal (E-CADHERIN), and connecting tubules (GATA-3) markers to show organoid patterning at D7 + 17 (scale bar, 200 μm). (D) and (E) Kidney organoids immunostained for podocyte (NEPHRIN) and parietal epithelial cell (CLAUDIN-1) markers showing similar podocyte maturity across cell lines and quantification at D7 + 17 (scale bar, 50 μm). Every dot represents a field of view.
Supplier Page from Abcam for Anti-NPHS2 antibody