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. 2015 Jan 20;290(11):6810–6824. doi: 10.1074/jbc.M114.606699

FIGURE 1.

FIGURE 1.

CXCR4-promoted lysosomal degradation of DEPTOR. A, CXCR4 activation induces degradation of DEPTOR. HeLa cells were treated with vehicle (PBS + 0.1% BSA) or CXCL12 for the indicated times, as described under “Experimental Procedures.” Equal amounts of whole cell lysates were immunoblotted for DEPTOR and the indicated proteins. B, immunoblots were analyzed by densitometry. Graph shows the average amount of DEPTOR remaining normalized to actin at each CXCL12 time point compared with vehicle (0 time point; set to 100%) from four independent experiments. Error bars represent the mean ± S.E. C, analysis of DEPTOR degradation in the presence of lysosomal or proteasomal inhibitors. HeLa cells were pretreated with vehicle (Veh), chloroquine, or lactacystin, followed by treatment with vehicle, CXCL12, or serum (10% FBS) for 3 h, as described under “Experimental Procedures.” Equal amounts of whole cell lysates were immunoblotted for DEPTOR and the indicated proteins. D, quantification of DEPTOR levels by densitometry. DEPTOR levels were normalized to actin, and the bars represent the average amount of DEPTOR remaining after CXCL12 or serum treatment compared with vehicle (set to 100%) under each inhibitor condition. Data are from 4 to 7 independent experiments, and error bars represent mean ± S.E. Data were analyzed by a two-way ANOVA followed by Tukey's post hoc test. *, data are significant from control (p < 0.05).