Article Figures & Data
Figures
- FIGURE 1.
Histologic changes in glioblastoma microenvironment 10 d after 225Ac-E4G10 treatment. (A) Confocal immunofluorescence imaging of tumor sections co-stained for anti-CD31, anti-desmin, and anti–collagen IV. Overlay of all stains is presented, with colocalization of endothelial cells (red) and pericytes (green) in yellow and counterstaining with DAPI (blue). Scale bars are 20 μm. (B and C) Quantification of pericyte density (desmin normalized by DAPI) (B) and pericyte coverage of blood vessels (desmin normalized by CD31) (C). (D) Immunohistochemical staining for regulatory T cells. Arrows indicate FoxP3-positive cells. (E) Quantification of FoxP3-positive regulatory T cells. Scale bars are 100 μm (full panels) and 20 μm (inset panels). Representative images of treatment and control groups are shown in A and D; positively stained area was counted and calculated as percentage of whole-tumor section for quantifications shown in B, C, and E. Data are mean ± SEM. HSA = human serum albumin.
- FIGURE 2.
Diffusion-weighted MRI study of vessel functionality after 225Ac-E4G10 treatment. (A) Representative MR images on day 10 after treatment: axial T2-weighted fast spin echo (left); corresponding echoplanar at b = 50 (predominantly perfusion) (middle); and echoplanar at b = 600 (predominantly diffusion) (right). Scale bars indicate relative intensities; dotted outline, tumor margin; solid arrow, tumor; and dashed arrows, water-filled phantom (measurement control). (B) Representative logarithmic plot of signal intensities of whole-tumor ROIs—plotted normalized to intensity at b = 0 and as natural logarithm (ln(Sb/Sb=0))—vs. measured b-values. Biexponential decay fitting of data (intravoxel incoherent motion, straight line) results in 2 slopes, D* (red line) and D (green line). ADC is derived by monoexponential decay fitting of data (dashed line). (C–E) Quantification of coefficients: ADC (C), D (D), and D* (E). Data are mean ± SEM.
- FIGURE 3.
Dasatinib penetration of glioblastomas 10 d after 225Ac-E4G10 treatment. Representative axial whole-brain sections stained with hematoxylin and eosin are shown, with matching autoradiography of 14C-labeled dasatinib from mice treated with vehicle (n = 5) or 225Ac-E4G10 (n = 6). Arrows indicate tumor margins. Scale bar indicates relative intensities. Autoradiography data are quantified on right. Data are mean ± SEM of mean intensities of whole-tumor ROIs. ***P < 0.001.
- FIGURE 4.
(A) Flow cytometry gating scheme to identify bone marrow EPCs in Ntva mice. After gating out debris (panel 1) and cell doublets based on forward scatter (panel 2), population of live (DAPI-negative) TER 119–negative and CD11b-negative cells is selected (third panel) and identified as VEGF receptor 2–positive, c-kit–positive EPCs (red square, panel 4). (B and C) Pretreatment (B) and posttreatment (C) comparisons of EPC population between Ntva mice and controls. Data are mean ± SEM. FSC = forward scatter; SSC = side scatter.
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