FIG 2.

MG132- and bortezomib-induced formation of the Degs1 ladder. Cells maintained in serum and grown to 70% confluence were treated with ABC294640 (25 μM) or SKi (10 μM) for 24 h. In certain cases, cells were treated with scrambled siRNA or Degs1 siRNA (each at a final concentration of 100 nM) for 48 h or with MG132 (10 μM) for 30 min prior to treatment with ABC294640 or SKi. Cells were also treated with bortezomib (5 or 10 nM) for 24 h. (A) Western blot probed with anti-Degs1 antibody showing the effect of Degs1 siRNA on Degs1 laddering induced by MG132 in the presence or absence of SKi or ABC294640. (B) Western blot probed with anti-Degs1 antibody showing the effect of bortezomib (BTZ) on Degs1 laddering. (C) Western blot showing the formation of the Degs1 ladder in response to SKi or MG132 in PANC1 cells. Blots were reprobed for GAPDH using anti-GAPDH antibody to ensure comparable protein loading. Results are representative of data from at least 3 independent experiments. Lanes C, control.