Figure 6.

Expression and subcellular localization of TORC2 in EL4-Gax cells. A. Immunofluorescent staining of TORC2 protein (red) in EL4-Gax cells grown in vitro, in vivo, or ex vivo. Cells were counterstained with DAPI (blue) to localize the nucleus and examined by confocal microscopy. The right panel shows a magnified image of a single cell indicated by an arrow in the adjacent panel. B. Statistical analysis of subcellular localization of TORC2 expression. The amount of protein expression was determined as the number of fluorescent pixels in the total and nucleus for each growth condition. Data were obtained by counting pixels in 108, 137, and 106 cells for in vitro, in vivo, and ex vivo conditions, respectively. Error bars indicate SEMs. *; p < 0.001. C. Expression of AMPKα protein in EL4-Gax cells. Anti-AMPKα and anti-phosphor-AMPKα (Thr172) antibodies were used to detect protein in EL4-Gax cells grown in vitro, in vivo, and ex vivo. Equivalent protein loading was confirmed by stripping and re-probing the blot with an anti-β-actin antibody.