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. Author manuscript; available in PMC: 2017 Nov 1.

Published in final edited form as: Biochim Biophys Acta. 2016 Sep 4;1861(11):1808–1815. doi: 10.1016/j.bbalip.2016.09.001

Treatment of serum-starved MCF-7 cells, which have very low constitutive phosphorylated Akt, with D-3-deoxy diC₈PI induces aoptosis. (A) Akt phosphorylation on Ser473 (pAkt Ser473) was measured in whole MCF-7 cell detergent lysates by western blot analysis. The blot was stripped and subsequently probed for constitutive expression of HSP90. The serum-starved MCF-7 cells were incubated with PBS (control), 100 μM D-3-deoxy-diC₈PI (D-3-deoxy), or 16 μM perifosine for 3.5 h. As a positive phosphorylation control, cells were incubated with PBS without serum for 3 h 10 min and then stimulated with 100 ng/mL HRG β1 for 20 min. (B) Viability curves for cells treated with D-3-deoxy diC₈PI (▴ ) or the alkylphospholipids miltefosine (●) and perifosine () for 24 h in the absence of serum. The inset shows the viability data for the two alkylphospholipids. (C) D-3-deoxy diC₈PI induces apoptosis in MCF-7 cells as evidenced by PARP cleavage after 6.5 h and caspase 9 cleavage after 3.5 h incubation.

Inline graphic — Treatment of serum-starved MCF-7 cells, which have very low constitutive phosphorylated Akt, with D-3-deoxy diC₈PI induces aoptosis. (A) Akt phosphorylation on Ser473 (pAkt Ser473) was measured in whole MCF-7 cell detergent lysates by western blot analysis. The blot was stripped and subsequently probed for constitutive expression of HSP90. The serum-starved MCF-7 cells were incubated with PBS (control), 100 μM D-3-deoxy-diC₈PI (D-3-deoxy), or 16 μM perifosine for 3.5 h. As a positive phosphorylation control, cells were incubated with PBS without serum for 3 h 10 min and then stimulated with 100 ng/mL HRG β1 for 20 min. (B) Viability curves for cells treated with D-3-deoxy diC₈PI (▴ ) or the alkylphospholipids miltefosine (●) and perifosine () for 24 h in the absence of serum. The inset shows the viability data for the two alkylphospholipids. (C) D-3-deoxy diC₈PI induces apoptosis in MCF-7 cells as evidenced by PARP cleavage after 6.5 h and caspase 9 cleavage after 3.5 h incubation.