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. Author manuscript; available in PMC: 2015 Dec 17.
Published in final edited form as: Int J Cancer Clin Res. 2015;2(3):1–6. doi: 10.23937/2378-3419/2/4/1023

Figure 3. DIF-3(+1) inactivates purified PAK1 in vitro.

Figure 3

A) MCF-7 PAK1 cells were serum-deprived, pre-treated with vehicle or 20 μM DIF-3 (+1) for 2 h, and stimulated with or without PRL (200 ng/ml, 20 min), E2 (1 nM, 30 min) or HRG (30 ng/ml, 15 min). Immunoprecipitated HA-PAK1 was subjected to the in vitro kinase assay as described in Figure 2. Data are presented as in Figure 2B). Purified GST- tagged PAK1 was subjected to in vitro kinase assay in the presence of DIF-3(+1) or THPH. The same membrane was probed with anti-PAK1 antibody for detection of GST-PAK1. Relative PAK1 kinase activity was then normalized by the amount PAK1 for each lane. The numbers at the bottom give the relative fold increase of 32P incorporation into PAK1.