Figure 5. Blockade of eHsp90 signaling impairs spheroidogenesis.

A. Schematic representation of treatment regimens followed for cells treated with either 1 uM NPGA, 100 nM SCH or 500 nM GSK343. Cells were separated into 3 treatment groups, and each group was placed into spheroid suspension at Day 0. The pre-treatment group was subjected to the indicated inhibitors for 3 days prior to the spheroid suspension assay. The post-treatment group was treated with the indicated inhibitors upon initial suspension into the spheroid assay. The continuous treatment group (Contin) was pre-treated for 3 days with the indicated inhibitors, and subsequently maintained under the specified conditions following initiation of spheroid culture. ARCaPE-eHsp90 B-D. and DU145-eHsp90 E-G. were subjected to the indicated inhibitors and evaluated under the 3 experimental conditions as outlined in A. The number of spheres generated by each treatment status was counted for each generation and is graphically depicted. The sphere treatments were all compared against the untreated (UT) control sample for each given passage. All statistics were performed using the Student's t-test for the treated versus the untreated control. * = p<0.05, ** p<0.01.