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. 2011 Sep 7;302(1):F103–F115. doi: 10.1152/ajprenal.00419.2011

Fig. 2.

Fig. 2.

Wtip knockdown impaired stress fiber formation and dysregulated focal adhesion maturation. A: actin stress fibers were visualized by rhodamine-phalloidin staining in control vector (shEMP; top) and in shWtip (bottom) and costained with anti-WTIP (FITC). B: focal adhesions were immunostained with anti-vinculin (TRITC) in shEMP and shWtip. C: focal adhesion maturation was assessed by anti-vinculin immunostaining in shEMP and shWtip cells. Quantification of focal adhesion size was determined as described in experimental procedures. The bar graph shows mean relative intensity units for vinculin (right). D: shEMP and shWtip cells were immunostained with anti-paxillin, a marker of focal contacts. The bar graph shows mean relative intensity units for paxillin (right). E: phosphotyrosine levels of focal adhesions in shEMP and shWtip cells were assessed by immunostaining with PY99 as an indicator of focal adhesion maturation. The bar graph shows mean relative intensity units for phosphotyrosine (right). F: RT-PCR of shEMP and shWtip cells for Wtip, Limd1, and Gapdh transcripts. G: immunoblot analysis of shEMP (control) and shWtip (knockdown) podocytes for Wtip, paxillin, vinculin, focal adhesion kinase (FAK), and tubulin. Leica Quantify software was used for quantification. Scale bars = 20 μm. *P < 0.05.