Fig. 1.

Induction of TJ formation by SP600125 in HaCaT cells. a On day 1, the upper, TJ protein ZO-1 (red) was immunostained in cells treated with (SP600125) and without (control) SP600125. The central portion of the photographs were line scanned (dotted lines) and the signal intensities were plotted. Scale bars, 50 μm. Lower left, total number of TJs on 600 μm scanned line (200 μm × 3 lines) from each cell image. Lower right, a representative confocal image of cells treated with SP600125. Images from upper (9 μm from culture dish) and lower (4.5 μm from culture dish) planes, and that of Z-stack were shown. Scale bars, 25 μm. b Western blot analyses of expression of Keratin 10 (K10) and TGase-1. Cells treated with SP600125 significantly down-regulated these differentiation markers. Experiments were repeated three times. c Relative cell viability was measured via a metabolic assay using Alamar-blue, as described previously (Rampersad 2012). SP600125 exhibited a cytostatic or toxic effect in all the six experiments