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. 2013 Dec 6;3(12):e167. doi: 10.1038/bcj.2013.64

Figure 2.

Figure 2

Effects of targeting HSP90 on monoclonal protein trafficking. Intracellular lambda light chain levels were measured via ELISA. Data are expressed as percentage of control (mean±s.d., n=3) and are representative of at least two independent experiments. (a) RPMI-8226 cells were incubated for 48 h in the presence or absence of 0.5 μM 17-AAG (AAG) in combination with either 10 μM lovastatin (Lov), 10 μM digeranyl bisphosphonate (DGBP) or 5 mM 3-PEHPC (3P). The * denotes P <0.05 per unpaired two-tailed t-test and compares treatments with and without 17-AAG. (b) RPMI-8226 cells were incubated for 12–48 h in the presence or absence of 10 μM lovastatin (Lov) and/or 0.5 μM 17-AAG (AAG). The * denotes P <0.05 per unpaired two-tailed t-test and compares lovastatin-treated cells to control cells. The ** denotes P <0.05 per unpaired two-tailed t-test and compares lovastatin+17-AAG- treated cells with lovastatin alone. (c) ALMC-2 cells were incubated for 48 h in the presence or absence of 10 μM lovastatin (Lov) in combination with 0.5 or 1 μM 17-AAG (AAG). The * denotes P <0.05 per unpaired two-tailed t-test and compares treatments with and without 17-AAG. (d) RPMI-8226 cells were nucleofected with HSP90α/β siRNA (HSP90) or scrambled siRNA (Scr). After 24 h, cells were treated in the presence or absence of 10 μM lovastatin (Lov) for an additional 48 h. The * denotes P <0.05 per unpaired two-tailed t-test and compares the transfected cells to the untransfected cells (control or lovastatin). (e) RPMI-8226 cells were incubated for 24 h in the presence or absence of 0.5 μM 17-AAG (AAG) and brefeldin A (BFA, 0.1 or 0.5 μM). The * denotes P <0.05 per unpaired two-tailed t-test and compares treatments with and without 17-AAG. (f) Addition of a proteasome inhibitor augments the effects of lovastatin and 17-AAG. RPMI-8226 cells were incubated for 48 h in the presence or absence of 10 μM lovastatin (Lov), 0.5 μM 17-AAG (AAG) and 2.5 nM bortezomib (Bor). The * denotes P <0.05 per unpaired two-tailed t-test and compares treatments with and without bortezomib.