Figure 6. Pharmacologic Inhibition of c-Abl Increases Accumulation of Ganetespib in Human RCC Tumors from Patients.
(A) Human RCC tumors from patients were stained with hematoxylin and eosin (H&E). Patients 1 and 2 (P1 and P2) diagnosed with clear cell, Patient 3 (P3) with papillary type II, and Patient 4 (tumors 4a, 4b, and 4c) with papillary type I RCC are shown. The scale bar represents 20 μm.
(B) Within 15 min of removal of tumors in (A) by radical nephrectomy, they were dissected into 3 mm3 pieces that were cultured in mediumcontaining 5 μM GNF-5 for 24 hr and then with 0.5 μM BODIPY FL-ganetespib (STA-12-9455) for an additional 6 hr. There were 107 cells that were isolated from the tumors and the efficacy of c-Abl inhibition was confirmed by immunoblotting of the lysates with anti-phospho Y89-c-Abl monoclonalantibody.
(C) Cells from (B) were stained with PI, fixed, and analyzed by flow cytometry.
(D) Percentage (%) geometric mean fluorescence intensity (MFI) of the RCC cells from (B) treated with GNF-5 and/or FL-ganetespib.
(E) Geometric mean fluorescence intensity (MFI) data from (C) were transformed to log10 values prior to statistical analysis. For each tumor treatment, a scatter dot plot with the mean and the error bars representing the SE of the mean (*p < 0.05) is shown.