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. Author manuscript; available in PMC: 2010 Sep 15.
Published in final edited form as: Cancer Res. 2009 Aug 25;69(18):7311–7319. doi: 10.1158/0008-5472.CAN-09-1077

Figure 4. Expression of mutant PIK3CA increased the sensitivity of SW1736 cells to perifosine and temsirolimus.

Figure 4

A) Stable expression of PIK3CA mutants H1047R or E545K in SW1736 cells. SW1736 cells were transfected with lentiviral vectors containing mutant PIK3CA or empty vectors and selected with blastomycin for 2 weeks. Stable cell pools were lysed and immunoblotted with antibody against V5 tag or p-Akt to detect the expression of exogenous PIK3CA mutant and p-Akt level. The antibody against β-actin was used for quality control in Western blotting. B) Quantitative illustration of relative p-Akt levels. Densitometry was performed to measure the density of the p-Akt and β-actin bands in Fig 4A. The relative p-Akt levels were obtained by dividing the p-Akt band density by the corresponding β-actin band density. C) Effects of perifosine (left panel) or temsirolimus (right panel) on the proliferation of SW1736 cells transfected with PIK3CA constructs. Cells were treated with perifosine or temsirolimus at the indicated concentrations for 5 days with the culture medium and the drug replenished every 24 h. MTT assay was used to measure cell proliferation and IC50 values were calculated according to the Reed-Muench method (21).