Fig. 6.

Ganetespib treatment inhibits tumor growth, metastasis, and HIF-1 target gene expression following orthotopic transplantation of MDA-MB-435 cells. SCID mice received a mammary fat pad injection of MDA-MB-435 cells followed by tail vein injection of vehicle (Veh) or ganetespib (Gan; 150 mg/kg/week) starting on day 7 after orthotopic injection and weekly thereafter. Tumor volume (a) and body weight (b) were determined twice weekly and final tumor weight was measured (c). Lung sections were stained with H&E (d) and the number of metastatic foci per lung section was determined (e). Metastatic burden was determined by qPCR of lung genomic DNA using primers specific for the human HK2 gene (f). CTCs in peripheral blood were determined by RT-qPCR assay of total cellular RNA using primers specific for human 18S rRNA (g). Total RNA isolated from primary tumors was analyzed by RT-qPCR using primers specific for human VEGF, ANGPTL4, SDF-1, P4HA2, MMP9, and RPL13A mRNA normalized to the levels of 18S rRNA. For all bar graphs, ** p < 0.01, *** p < 0.001 vs vehicle, two-way ANOVA or Student’s t test (mean ± SEM; n = 6).