PKC412 induces modulation of c-fos and c-jun and increases AP-1 transcriptional activity. (A) RT-PCR of C-FOS following 90-minute pretreatment with PKC412 (10 μM) or DMSO (1:1000) followed by 30 minutes of PMA (100 nM) stimulation. (▪ indicates LP-1; ⊡, OPM2). Graph represents data from 2 individual experiments. Representative image shown. (B) RT-PCR of C-FOS following PKC412 (10 μM) for 30 minutes, 4 hours, and 20 hours (⊡ indicates NCI-H929). Graph represents data from 2 individual experiments. Representative image shown. For both panels A and B, Scion Image (Scion, Frederick, MD) was used for densitometric analysis. (C) Reduction in nuclear expression of c-fos after 24 hours of treatment with 2 μM and 4 μM PKC412 was determined by Western blot. (D) RT-PCR of C-JUN after PKC412 (10 μM) treatment for 30 minutes, 4 hours, and 20 hours (⊡ indicates NCI-H929). Graph represents data from 2 individual experiments. Representative image is shown. (E) PKC412 (P) (5 μM) treatment induces transcriptional activation of AP-1 as determined by luciferase assay (measured after 18 hours of treatment) (⊡ indicates NCI-H929). Graph represents results from 2 experiments. Error bars for all graphs indicate SEM.