Figure EV1. mTOR remains at the lysosome in starved KO myotubes, but not in myoblasts.

1. Lysosomal localization of mTOR after starvation (HBSS) in KO cells was confirmed by using a different immortalized KO cell line (clone RF1). The cells were grown in differentiation medium for 8 days, starved for 2 h, and fixed. The images show immunostaining of fed (NT) and starved (HBSS) KO cells with anti‐LAMP1 (red) and anti‐mTOR (green) antibodies. Scale bar: 10 μm.
2. Immunostaining of KO myoblasts grown in proliferation medium with anti‐LAMP1 (red) and anti‐mTOR (green) antibodies. Lysosomal localization of mTOR is observed in fully fed KO myoblasts (NT). Almost no co‐localization of the two stains is seen in KO myoblasts after 2 h of starvation (HBSS). The images show randomly selected fields of > 100 cells for each condition. Scale bar: 10 μm.