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. 2011 Nov 30;6(1):33–47. doi: 10.1016/j.molonc.2011.11.008

Figure 3.

Figure 3

shRNA against uPA and uPAR down regulates phosphorylation of JAK2/STAT5 in glioblastoma and endothelial cells and phosphorylation of GM‐CSFrβ at Tyr 766 position in HMEC, U87MG and 4910 cells. (A) Western blot analysis of shRNA‐transfected HMEC, U87MG, and 4910 cells was carried out with primary antibodies against pJAK2 (Tyr1007/1008) and pSTAT5 (pTyr 695/699) and with total forms of JAK2 and STAT5. A representative blot of three independent experiments is shown. (B and C) Quantitative analysis was done for pJAK2 and pSTAT5 by optical densitometry as per standard protocols. (D) Western blot analysis of shRNA‐transfected HMEC, U87MG, and 4910 cells was carried out with primary antibody against pTyr766 position of the GM‐CSFrβ unit and appropriate secondary antibody. GM‐CSFRβ antibody was also immunoblotted to demonstrate equal loading of lysates. A representative blot of three independent experiments is shown. (E) Quantitative analysis was done by optical densitometry as per standard protocols.