Fig. 6

Src overexpression rescues cell proliferation and adhesion of plectin knockout melanoma cells. A Cells were plated at a density of 2.5 × 10⁴ cells per well in 24-well plates and the number of cells quantified with an automated cell counter (n = 6). B Cells were plated at 1 d after Src introduction. The number of viable cells were determined by CCK-8 assay at indicated time (n = 3). C Cells were fixed and stained with DAPI, anti-Ki-67/Alexa fluor 555 and anti-Flag/Alexa fluor 488 at 1 d after plating. The percentage of Ki-67 positive cells relative to DAPI was calculated. Scale bur = 100 μm (n = 5). D Src was introduced into the cells for 1 d. Cells were cultured at 5 × 10³ cells per 15 μl drop on a dish lid to form spheroid for 7 d. Spheroids were observed by stereoscopic microscope and measured by ImageJ. Scale bar = 500 μm (n = 6). E The confluent cells were detached with dispase and number of cell fragments was taken photo (left panel) and quantified (right panel). Scale bar = 20 mm (n = 6). F Cells were plated on a fibronectin-coated dish 1 d after Src introduction. After 2 h incubation, the adherent cells were stained with crystal violet (left 4 panels) and counted (right panel). Scale bar = 10 μm (n = 6), *; p < 0.05 vs control Src(−), #; p < 0.05 vs PKO Src(−), ns; no significance vs control Src(−)