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

FIGURE 2.

FIGURE 2.

Depletion of ESCRT-I subunit UBAP1 attenuates CXCR4 and DEPTOR degradation. A–F, HeLa cells treated with siRNA directed against UBAP1, Mvb12A, Mvb12B, or luciferase (Ctrl) were stimulated with vehicle (minus symbol) or CXCL12 (plus symbol) for 3 h, as described under “Experimental Procedures.” Equal amounts of whole cell lysates were immunoblotted for CXCR4 (A and C) or DEPTOR (E) and the indicated proteins. Bars represent the average amount of CXCR4 degraded compared with vehicle control (B and D) or DEPTOR remaining (F) after each treatment condition, compared with vehicle and luciferase siRNA-transfected cells (set to 100%). CXCR4 (B and D) and DEPTOR (F) levels were normalized to actin. Data are from 3 (B and D) or 4 (F) independent experiments. Error bars (B, D, and F) represent mean ± S.E. Data were analyzed by a Student's t test (B), one-way ANOVA (D), or two-way ANOVA (F). CXCR4 degraded in UBAP1 siRNA-treated cells is significantly different from control cells +(p = 0.003).