Effect of exogenous IGF-1 on expression of TAP-1, pStat1(Tyr701) and pStat3(Tyr705). A,

<p>Expression of TAP-1 peptide in T98G cells was down-regulated by addition of IGF-1 to culture medium. TX cells were incubated in absence of serum and in the presence of 50 μM ZnSO₄ and IGF-1 (100 ng/ml) over-night. Lanes 1 and 2 are NT cells from the T98G and HG-2 cell lines respectively. Lane 3 and lane 4 are T98G TX (clone b) and HG-2 TX (clone 5), respectively, into which IGF-1 was added overnight. Lanes 5 and 6 are as in lanes 3 and 4 except without added IGF-1. <b>B.</b> The expression of TAP-1 was examined as a function of time following addition of IGF-1 (100 ng/ml) in TX T98G cells in absence of serum and in the presence of Zn2+. Cells were treated with exogenous IGF-1 for the time periods as depicted. The cell lysates were prepared and subjected to Western Blot analysis as described in methods. The primary antibodies to TAP-1, pStat1 (Tyr701), pStat3 (Tyr705) and Stat3 were also obtained as described. The 70 kd TAP-1, 91 kd pStat1, and 86 kd pStat3 and 86 kd Stat3 peptides produced during the time course were identified by monoclonal antibody used and molecular weights. The data represent one of three sets of similar experimental results. <b>C,</b> Data from <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0058428#pone-0058428-g007" target="_blank"><b>Fig 7</b></a><b>B</b> and similar results from two repeat experiments were used for this bar graph. The densitometry quantification was done using the NIH image J program as described in methods.