Effect of TPA treatment on desmosome and protein solubility in A431 keratinocytes.

<p><b>(A)</b> Parental and <i>KRT17</i> null A431 cells were stained for desmoplakin, E-cadherin, and nuclei (DAPI) 30 min after acetone-control Tx, or 30 and 60 min after TPA Tx. Scale bars: 10 μm. Arrows point to desmoplakin staining at cell–cell borders, and arrows with asterisks denote partial loss of desmoplakin staining at cell–cell borders after TPA Tx. <b>(B, C)</b> Quantification of desmoplakin and E-cadherin staining at cell boundary (ROI width = 1 μm, see Methods). Each dot represents a cell. Data are shown as mean ± SEM. Two-way ANOVA. <b>(D)</b> Western blot analysis of the solubility of K17, PKCα, RACK1, and (control) β-Actin in parental A431 cells 1 h after TPA Tx, and 20 μg of total protein was loaded in each lane for the low-salt and triton-soluble fractions, and 2 μg of total protein was loaded in each lane for the triton-insoluble fractions. The source data used to derive the numerical values reported here can be found in <b><a href="http://www.plosbiology.org/article/info:doi/10.1371/journal.pbio.3002779#pbio.3002779.s016" target="_blank">S1 Data</a></b>.</p> <p>(PDF)</p>