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. 2005 Jul;7(7):638–645. doi: 10.1593/neo.04736

Figure 2.

Figure 2

Decrease in endogenous PAK1, Akt, and BAD phosphorylation in MCF10A cells grown in suspension. MCF10A cells were grown either attached (t = 0) or suspended for 2, 4, or 8 hours. (A) Western blots of whole cell lysates were probed with anti-phospho Akt (Ser-423) to detect active Akt, and then stripped and reprobed with anti-Akt to detect total Akt. (B) Western blots of whole cells lysates were probed with anti-phospho PAK1 (Thr-423) to detect active PAK1 (the doublet observed may be due to the fact that the antibody will also react with any endogenous PAK2 that may be present and phosphorylated on residue Thr-402), and then stripped and reprobed with anti-Pak c-19 to detect total PAK1. (C) Western blots of whole cell lysates were probed with anti-phospho-BAD (Ser-136) to detect phosphorylated BAD, then stripped and reprobed with anti-phospho-BAD (Ser-112) to detect phosphorylated BAD, and then stripped and reprobed with anti-BAD to detect total BAD.