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. Author manuscript; available in PMC: 2014 Jul 1.
Published in final edited form as: Cancer Discov. 2013 Apr 4;3(7):782–797. doi: 10.1158/2159-8290.CD-12-0215

Figure 5. DGKα attenuation is safe in normal cells.

Figure 5

A, Normal human astrocytes were transfected with either control or DGKα siRNA, and cell number was assessed at 3 days post transfection. B, An immunoblot was done on cell lysates post-transfection to check for transfection efficiency. C and D, Normal human astrocytes and fibroblasts were treated with 10 µM R59022, R59949, or DMSO (v:v) control and cell proliferation was assessed at 3 days post treatment, with no significant decrease observed in cell number. E, Levels of cAMP were also evaluated via ELISA 5 days after knockdown and inhibition of DGKα in astrocytes. F, Effects of exogenous cAMP on mTOR transcription in astrocytes were assessed through mTOR promoter luciferase assay 5 days after treatment. G, Rolipram was administered at 40 µM to further test the effect of phosphodiesterase inhibition/cAMP levels on mTOR transcription in astrocytes, with promoter activity assayed at 6 days post treatment. (*, P<0.05 and **, P<0.01 Student t test)