Figure 3: Adult CLIC4-KO PTs have dilated lumens and abnormal MV morphogenesis. | Nature Communications

Figure 3: Adult CLIC4-KO PTs have dilated lumens and abnormal MV morphogenesis.

From: CLIC4 regulates apical exocytosis and renal tube luminogenesis through retromer- and actin-mediated endocytic trafficking

Figure 3

(a,b) PAS staining of 3-week-old WT (a) and CLIC4-KO (b) renal cortical sections. An arrow in (a) points to strong PAS-labelled brush borders facing a PT lumen. (c,d) EM analysis of 3-week-old WT (c) and CLIC4-KO (d) mouse PT. Red brackets indicate the RE-containing apical cytoplasmic regions. Arrows in (d) point to the large vacuoles in the mutant PT. Inset in (d) shows a low-magnification view of a dilated PT that failed to develop a typical MV-lining lumen. (e,f) Megalin immunostaining of adult WT (e) and mutant (f) PTs. Megalin had a lumen outlining pattern (arrows) in WT. Mutant PTs displayed weak megalin signals (arrowheads). (g) Immunoblots of lysates of LLC-PK1(CL4) cells without (WT) or with (KD) doxycycline-induced CLIC4-shRNA expression. (h,i) LLC-PK1(CL4) cells, without (h) or with (i) expression of CLIC4-shRNA/RFP, stained with megalin and β-catenin. Representative confocal images of x-y (upper panel) and x-z (two lower panels; arrows point to the apical side of cells) views are shown. Note that in (i) the ‘red’ CLIC4-shRNA-expressing cells had selectively reduced megalin signal. Scale bars, 100 μm (a,b); 2 μm (c,d); 50 μm (e,f); 10 μm (h,i).

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