Fig. 4
Impact of PI3K/mTOR inhibition in PTEN deficient cells. A Representative Immunoblot of KP6 and KPP4 cells without and with 2 µM PI-103 treatment 3 h before irradiation with 8 Gy at time points directly, 2 h, 4 h, 8 h and 24 h after irradiation. DMSO as solvent control. Actin and AKT as loading control. n = 3. B Colony formation assay KP6 (black) and KPP4 (blue) cells with 2 µM PI-103 (dashed lines) and DMSO as control (continuous lines) with re-seeding protocol (Additional file 1: Fig. S4A). SF 2: Surviving fraction at 2 Gy. D25: Dose in Gy with 25% survival. Error bars: Standard deviation. n = 3. C Relative number of living of KP6 (black) and KPP4 (blue) cells 27 h after treatment with PI-103 in different concentrations, DMSO as control and 24 h after radiation with 8 Gy (dashed lines) or without radiation (continuous lines) (dead cells stained with trypan blue excluded from analysis). Error bars: Standard deviation. n = 3. D Immunoblot of (phospho-)AKT and (phospho-)S6 BEAS-2B wildtype (WT), PTENhomo, BRAFV600E and compound mutant cell lines without and with 2 µM PI-103 pre-treatment for 3 h. DMSO as solvent control. Actin serves as loading control. E Colony formation assay of WT (black) and PTEN deficient (blue) BEAS-2B cells with 3 h pre-treatment of 2 µM PI-103 (dashed lines) and DMSO as control (continuous lines) with 24 h re-seeding protocol (Additional file 1: Fig. S4A). SF 2: Surviving fraction at 2 Gy. D25: Dose in Gy with 25% survival. Error bars: Standard deviation. n = 3. Also see Additional file 1: Fig. S4