Journal: PLoS ONE

Article Title: Characterization of a Human Cell Line Stably Over-Expressing the Candidate Oncogene, Dual Specificity Phosphatase 12

doi: 10.1371/journal.pone.0018677

Figure Lengend Snippet: A. Left: GFP or GFP-DUSP12 cells were treated with DMSO or 5 µM staurosporine (STS) overnight and the Promega Caspase 3/7 Glo assay was used to measure apoptosis. Means of three independent experiments are graphed with error bars representing SEM. Significance of a P value<0.05. Right: Immunoblot detecting cleaved PARP in lysates of GFP and GFP-DUSP12 cells treated overnight with 5 µM STS. Lysates were collected with RIPA buffer and equalized by total protein using a BCA assay. A representative blot from three independent experiments is shown. B. Left: GFP or GFP-DUSP12 cells were treated with DMSO or 200 nM thapsigargin (TG) for 48 hours and the Promega Caspase 3/7 Glo assay was used to measure Caspase 3/7 activation as an indication of apoptosis. Means of three independent experiments are graphed with error bars representing SEM. Significance of a P value<0.05. Right: Immunoblot detecting cleaved PARP in lysates of GFP and GFP-DUSP 12 cells treated with 200 nM TG for 48 hours. Lysates were collected with RIPA buffer and equalized by total protein using a BCA assay. Results depicted are representative of at least three independent experiments.

Article Snippet: The next day, either staruospoine (STS) or thapsigargin (TG) (Acros Organics) were added to the cells at a final concentration of 5 µM STS or 200 nM TG, and incubated for an additional 16 hours or 48 hours respectively.

Techniques: Glo Assay, Western Blot, BIA-KA, Activation Assay