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. 2014 Feb 19;289(14):9502–9518. doi: 10.1074/jbc.M113.505743

FIGURE 5.

FIGURE 5.

TAT-Neph1CD-transduced podocytes are resistant to PAN-induced cytoskeletal damage. A and B, TAT-GFP- and TAT-Neph1CD-transduced podocytes were treated with PAN for indicated time periods (0–48 h) and immunostained with phalloidin (Alexa 488) and Neph1 (Alexa 594), and mounted with DAPI. Confocal imaging showed that PAN treatment resulted in drastic changes to the actin cytoskeleton in TAT-GFP-transduced cells. Additionally, the distribution of junctional proteins, including Neph1, was significantly altered in these cells. In contrast, the PAN treatment of podocytes transduced with TAT-Neph1CD showed minimal alteration to the actin cytoskeleton with increased localization of Neph1 at the junctions. Confocal images were collected at ×60 magnification and presented after deconvolution in XY and XZ orientation. B, pixel intensity profile was created by placing a line spanning the cell-cell junction and analyzed using ImageJ software. This analysis suggested that the intensity of Neph1 at the cell-cell junctions was maintained in TAT-Neph1CD-transduced cells treated with PAN (p < 0.05). Scale bar, 10 μm (A).