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. 2003 Mar 31;160(7):1001–1008. doi: 10.1083/jcb.200212113

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Copyright © 2003, The Rockefeller University Press

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Figure 3. — CH-ILKBP regulates PKB/Akt phosphorylation. (A and B) The CH-ILKBP siRNA transfectants (lanes 1 and 4), the control RNA transfectants (lanes 2 and 5), and the parental HeLa cells (lanes 3 and 6) were serum starved (lanes 1–3) and stimulated with 10% FCS (A, lanes 4–6) or 2 ng/ml IGF-1 (B, lanes 4–6) for 10 min. The cell lysates (10 μg/lane) were analyzed by Western blotting with antibodies recognizing Akt, phospho-Akt(Ser473), phospho-Akt(Thr308), phosphor-PDK1(Ser241), and PDK1, respectively. (C) The CH-ILKBP siRNA transfectants (lanes 1 and 2), the control RNA transfectants (lanes 3 and 4), and HeLa cells (lanes 5 and 6) were kept in suspension for 2 h and then either harvested (lanes 1, 3, and 5) or allowed to adhere on laminin-coated plates for 30 min (lanes 2, 4, and 6). The cell lysates (15 μg/lane) were analyzed by Western blotting with antibodies recognizing Akt, phospho-Akt(Ser473), and phospho-Akt(Thr308), respectively.