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2 protocols using p stat3 p y705

1

STAT3 Immunoblotting Protocol

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Cells were harvested, lysed in RIPA buffer (Santa Cruz Biotechnology; Santa Cruz, CA), and protein was quantified and electrophoresed as previously described (7 (link)). Immunoblotting was performed using the following antibodies: total STAT3, and p-STAT3(p-Y705) (Cell Signaling; Danvers, MA) at a 1:1000 dilution.
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2

Sorafenib and Everolimus Combination Therapy

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Sorafenib tysolate and everolimus were purchased form L.C. Laboratories (Woburn, MA). For all in vivo studies, sorafenib and everolimus were prepared as previously described [9 (link),30 (link)]. Antibodies for western blot and immunohistochemical analysis were purchased as follows: β-catenin (#8480), Bcl-2 (#2870), BIM (#2933), cleaved caspase-3 (#9661), ERK (#9102), GAPDH (#2118), mTOR (#2983), p-β-catenin (#9561), p-ERK (#4370), p-mTOR (#2971), p-S6 (#2211), p-STAT3-pY705 (#9145), p-STAT3-pY727 (#9134), S6 (#2217) and STAT3 (#4904) antibodies from Cell Signaling Technology (Danvers, MA); AR (#RB-9030), PCNA (#RB-9055) and Ki67 (#RB-9043) antibodies from Thermo Scientific (Waltham, MA); cyclin B1 (sc-752), cyclin D1 (sc-753) antibodies from Santa Cruz Biotechnology (Dallas, TX); CD31 (#550300) from BD Biosciences (San Jose, CA).
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