Figure 6
From: Activity-independent targeting of mTOR to lysosomes in primary osteoclasts
mTOR remains associated with the lysosomes in osteoclasts during amino acid starvation. (A) RAW264.7-derived osteoclasts (RAW-OCs) were differentiated for 4 days, incubated with dextran coated magnetic nanobeads for 24 hrs, chased for additional 24 hrs, and lysosomes purified as described in “Materials and Methods”. (B) Purified lysosomes from the undifferentiated RAW264.7 cells were isolated as described in “Materials and Methods”. Equal amounts of protein were loaded per lane and the blots were probed for mTOR, p-mTOR, Raptor, LAMTOR1, LAMP2, and the V-ATPase subunits a3 and A. Representative cropped blots of three independent experiments for RAW-OCs and for undifferentiated RAW264.7 cells. CTRL = control, no treatment; STRV = starvation, HBSS, 1 hr.
