Quantity one analysis software program

Total cell lysates, cytoplasmic and nuclear protein fractions were extracted as described previously (20 (link)). Standard Western blotting was carried out using primary anti-Merlin, anti-FOXM1, -β-catenin, -cyclin D1, -c-Myc, -μPAR, -MMP9, and -VEGF (rabbit; Santa Cruz Biotechnology), and anti-MMP2 (rabbit; Cell Signaling Technology) antibodies. Equal protein-sample loading was monitored using an anti-GAPDH antibody for total cell protein lysates (rabbit; Santa Cruz Biotechnology), an anti-α-tubulin antibody for cytoplasmic fractions (mouse; Oncogene), or an anti-histone H1 antibody for nuclear fractions (mouse; Santa Cruz Biotechnology). Secondary antibodies were anti-mouse IgG or anti-rabbit IgG (Santa Cruz Biotechnology). The bands were quantified using the Quantity One analysis software program (version 4.6; Bio-Rad).

Slot blot analysis was expressed as the mean ± the standard deviation for three independent experiments from separate harvests. Relative Pt-damage was calculated by manual analysis of slot blot data using Quantity One^® analysis software program (Bio-Rad). Analysis of western blots used densitometry normalized to actin. Data are the mean ± standard error of the mean. Statistical significance between groups (p< 0.05) was determined using ANOVA with Tukey’s post hoc test or Student’s t-test as indicated.