Four-micron sections of formalin-fixed paraffin embedded organoids were stained with hematoxylin and eosin (Ventana Medical Systems, Tucson, AZ). Immunohistochemistry was performed with an automated, validated, and accredited staining system (Ventana Benchmark ULTRA) using the UltraView or OptiView universal DAB detection Kit (Ventana Medical Systems). After deparaffinization and heat-induced antigen retrieval, the tissue samples were incubated with WT1 (Cell Marque, Rocklin, CA), ECAD (Ventana Medical Systems), Villin-1 (Abcam, Cambridge, UK), CD31 (Cell Marque), renin (Abcam), NKCC2 (StressMarq Biosciences, Victoria, BC, Canada), NHE3 (StressMarq), CD34 (Cell Marque), COL1A1 (Novus Biologicals, Littleton, CO), or PDGFRa (R&D systems). Incubation was followed by hematoxylin II counterstaining. Double stainings of PDGFRa and renin were performed by incubating the samples with PDGFRa for 32 minutes at 37 C followed by detection with Red610 kit (#760-245, Ventana) and renin for 32 minutes at 37 C followed by detection with FAM kit (#760-243, Ventana). An AF488-conjugated anti-human mitochondria antibody MAB1273A4 (Millipore, Billerica, MA) was used to detect
Shankar A.S., Du Z., Mora H.T., Bosch T.v.d., Korevaar S.S., Garrelds I.M., Bresnick E., Iglesias C., Groningen M.C.v., Gribnau J., Baan C.C., Danser J., Hoorn E.J., & Hoogduijn M.J. (2020). Human kidney organoids produce functional renin. Cytotherapy, 22(5), S10-S11.
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