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. 2009 Nov 20;4(11):e7919. doi: 10.1371/journal.pone.0007919

Figure 3. The PI3K/Akt pathway regulates Fas expression and cell death during F. tularensis subsp. novicida infection.

Figure 3

(A) RAW 264.7 cells were pretreated with PI3K inhibitor LY 294002 for 30 minutes and then infected. Lysates were analyzed by Western blotting with Fas antibody. The same membranes were reprobed with actin antibody. The graph shows mean and SD of band intensity values obtained from three independent experiments. Data were analyzed by a student's t-test. * indicates p value <0.05. (B) Cell lysates were analyzed with phospho-Akt antibody to verify inhibition of the PI3K pathway. (C) Cell death was analyzed by measuring the amount of LDH release. (D) RAW 264.7 cells were transfected with empty vector or plasmids expressing wild-type Akt. The transfectants were infected and Fas protein expression was determined by Western blotting. The graph shows mean and SD of Fas band intensities from three independent experiments. * indicates p value <0.05. (E) Cell death was assessed by LDH assay. The graph shows mean and SD of LDH release from three independent infections. (F) Over-expression was confirmed by Western blot analysis with Akt antibody.