Fig. 1.

TORC1 and TORC2 are expressed in adult mouse brain and translocate to the nucleus of cultured neurons upon activation of calcium and cAMP pathways. (A) TORC1 and TORC2 mRNA levels in mouse neuronal cultures and adult brain as measured by real-time RT-PCR. Cx, cortex; Hipp., hippocampus; Str., striatum. (B) Nuclear translocation of endogenous TORCs in mouse cortical neurons exposed to FSK and KCl (preincubation with LMB). Microtubule-associated protein 2 (MAP-2) and DAPI stain neuronal processes and nuclei, respectively, are shown. (C) Immunofluorescence of neurons transfected with mouse TORC1-myc showing its nuclear translocation only in the presence of FSK and KCl (preincubation with LMB). (D) Immunofluorescence of neurons transfected with mouse Flag-TORC2 showing its nuclear translocation in the presence of FSK and KCl (no preincubation with LMB). Inhibition of calcineurin by CsA blocks the nuclear accumulation of TORC2 triggered by FSK and KCl. DAPI staining is shown to localize nuclei.