Targeting Leukemia-Associated Hypoxia with PR-104 Figure 8. Dose-response efficacy of PR-104 against ALL-8 and ALL-19 in vivo. (A and C) The responses of NOD/SCID mice engrafted with ALL-8 (A) or ALL-19 (C) xenograft to treatment with PR-104 at decreasing doses are represented as percentages of human CD45+ cells in the peripheral blood of individual mice over time. PR-104 was administered once a week for 6 weeks via i.p. injection; the shaded area corresponds to the duration of treatment. Kaplan-Meier plots of EFS, from which LGD was calculated (difference between median EFS of treated and control mice) and compared using the logrank test. The corresponding values are shown in Table S1. (B and D) Distribution of objective response measures for ALL-8 (B) or ALL-19 (D) engrafted individual mice at each dose of PR-104 tested. MCR, maintained complete response; SD, stable disease; PD, progressive disease. doi:10.1371/journal.pone.0023108.g008 Aldrich. Secondary antibodies IRDye 800CW Donkey anti-mouse and IRDye 680CW Donkey anti-mouse were obtained from LI- COR Biosciences. The following antibodies were used for IHC analysis: anti-GFP (Santa Cruz Biotechnology), anti-HIF-1a (BD Biosciences), FITC-conjugated mouse monoclonal anti-PIM (HPI, Inc), anti-CD45 (Cell Signaling Technology), anti-CD22 (Novo- castra), anti-CXCR4 (Abcam), anti-SDF-1a, anti-CD31 (Santa Cruz Biotechnology) and anti-CAIX (Novus Biologicals). HRP conjugated secondary antibodies were from Dako (LSAB+sys- tem+HRP). PLoS ONE | www.plosone.org 10 August 2011 | Volume 6 | Issue 8 | e23108