Expression of HSPs by 12B1-D1 cells.

(A) Surface expression of HSP72 and HSP60 on 12B1-D1 cells as determined by flow cytometry. Panel 1: 12B1-D1 cells were incubated with isotype control antibody followed by PE-conjugated goat antimouse IgG and analyzed by flow cytometry. Panels 2-5: 12B1-D1 cells incubated with anti-HSP72 antibody (left) and anti-HSP60 antibody (right) followed by PE-conjugated goat antimouse IgG. Panel 2: Untreated 12B1-D1 cells. Panel 3: 12B1-D1 cells were heat shocked for 1 hour at 42°C and then incubated for 6 hours at 37°C. Panel 4: 12B1-D1 cells were incubated for 6 hours with 40 nM AP20187. Panel 5: 12B1-D1 cells were heat shocked for 1 hour at 42°C, then incubated for 6 hours with 40 nM AP20187. (B) Immunoblot of 12B1-D1 cell lysates. Lane 1, untreated 12B1-D1; lane 2, 12B1-D1 + HS (A), cells were heat shocked at 42°C for 1 hour; lane 3, 12B1-D1 + HS (B), cells were heat shocked at 42°C for 1 hour and then incubated for 6 hours at 37°C; lane 4, 12B1-D1 + AP20187 for 6 hours; lane 5, 12B1-D1 + HS for 1 hour + AP20187 for 6 hours.