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. 2013 Nov 25;123(12):5179–5189. doi: 10.1172/JCI69000

Figure 3. ADCK4 localizes to the mitochondria and cytoplasm of podocytes in adult rat glomeruli and also in cultured human podocytes.

Figure 3

(A–C) Coimmunofluorescence of ADCK4 with (A) WT1 as well as (B) MTCO1 and (C) COXIV in adult rat glomeruli. ADCK4 partially colocalizes to mitochondria with the 2 mitochondrial markers COXIV and MTCO1. Scale bar: 10 μm. (D) Immunogold electron microscopy of adult rat kidney displays localization of ADCK4 (black arrowheads) at podocyte foot processes of glomeruli. A control without a primary antibody is shown on the left. Scale bar: 1 μm; ×2.5 (inset). (E) Podocytes were transfected with ADCK4-RFP and stained with an anti-COXIV antibody. ADCK4 partially colocalizes to mitochondria with COXIV. Note that ADCK4 also localizes along the plasma membrane (white arrowheads) in podocytes. Scale bar: 25 μm. (F) Subcellular fractionation of ADCK4 in undifferentiated and differentiated podocytes. Mitochondrial and cytosol fractions were prepared and immunoblotted for ADCK4, MTCO1, and COXIV, respectively. Each lane was loaded with 50 μg protein. Note that ADCK4 is present in both mitochondrial (marked by MTCO1 and COXIV), and cytosolic fractions in both undifferentiated and differentiated podocytes. W, whole cell lysates; Cyto, cytosol fraction; MT, mitochondrial fraction.