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Fig. 5 | Molecular Brain

Fig. 5

From: A role for BDNF- and NMDAR-induced lysosomal recruitment of mTORC1 in the regulation of neuronal mTORC1 activity

Fig. 5

Targeting mTOR to late endocytic compartments with photosensitive dimerizers. A Schematic representation of the optogenetic modules used to recruit mTOR on LEs. B Epifluorescence images illustrating the intracellular distribution of CIBN-myc-Rab7 on LAMP1 positive structures. Hippocampal neurons were transfected with CIBN-myc-Rab7 and immunolabeled for LAMP1 and myc. Bar, 10 µm. C DIV14 neurons were transfected with CIBN-myc-Rab7 and CRYII-mCherry-Raptor; two days later, cells were either kept in the dark (top images) or photoactivated for 5 min (15 × 100 ms-pulses of blue light at 2.42 mW/cm2; bottom images) before being processed for mTOR immunofluorescence. Upon photoactivation CRYII-mCherry-Raptor and endogenous mTOR are targeted to CIBN-myc-Rab7 positive structures. Boxed regions are shown at higher magnification D When CRYII-mCherry-Raptor was replaced with CRYII-mCherry, photoactivation induced the clustering of CRYII-mCherry on LEs but mTOR remained in the cytosol, as expected. Bar, 10 µm

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