Image-Guided Radiotherapy Targets Macromolecules. through Altering the Tumor Microenvironment
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1 Supporting Information Image-Guided Radiotherapy Targets Macromolecules through ltering the Tumor Microenvironment uthors: Oliver K. ppelbe,, Qingbei Zhang,, harles. Pelizzari, Ralph R. Weichselbaum,, and Stephen J. Kron,, ffiliations: Ludwig enter for Metastasis Research, The University of hicago, 5758 South Maryland venue, M 9006, hicago, IL 60637, United States Department of Molecular Genetics and ellular Biology, The University of hicago, 929 East 57 th Street, GIS W519, hicago, IL 60637, United States Department of Radiation and ellular Oncology, The University of hicago, 5758 South Maryland venue, M 9006, hicago, IL 60637, United States Stephen J. Kron, The University of hicago, GIS W522, 929 East 57 th Street, hicago, IL 60637, phone - (773) , fax - (773) , skron@uchicago.edu 1
2 0 Gy 8 Gy 1 Supplementary Figure 1. Radiation and perivascular apoptosis. Immunohistochemistry of tissue sections of MF7 GFP-IBD xenograft tumors excised 3 days after irradiation to detect cleaved caspase 3 (brown) displays no marked increase in apoptosis. Scale bar = 200 µm. 2
3 VEGFR-2 b only VEGFR-2 b+ B 15 VEGFR-2 b only VEGFR-2 b+ D31 %%area area 10 ^ α SM 5 ^ IV n M ol la ge αs VEGFR-2 b only VEGFR-2 b da 1 hr s y Radiant Efficiency (x 109) / tumor volume (mm3) ollagen IV D 31 0 Supplementary Figure 2. VEGFR-2 antibody depletes the tumor endothelium to the detriment of the radiation-enhanced delivery effect. () Immunohistochemistry of MF7GFP-IBD xenograft tumors depicting the decrease in endothelium (D31, brown), pericytes (α-sm, brown), and basal lamina (ollagen IV, brown) following treatment with VEGFR-2 antibody. Purple = hematoxylin, nuclei. Scale bar = 200 µm. (B) 3
4 Relative quantification of IH staining in MF7 GFP-IBD tumor sections. % area denotes the area of an image stained calculated using an ImageJ macro (details in methods). p 0.05 relative to control, p 0.05 relative to only, ^ p 0.05 relative to VEGFR-2 antibody only, n = 3. () MF7 GFP-IBD tumor retention of ngiosense, measured using IVIS fluorescence quantification, appears limited by the stromal changes apparent following VEGFR-2 treatment. ngiosense was administered 3 days after IR. p 0.05 relative to control at the same time point, n = 3. 4
5 imatinib only imatinib+ B 15 D31 imatinib only imatinib+ % % area area 10 α SM 5 Radiant Efficiency (x 109) / tumor volume (mm3) IV n M ol la ge αs ollagen IV D imatinib only imatinib ^ Supplementary Figure 3. Imatinib mesylate treatment leads to depletion of pericytes but does not act synergistically with IR to improve radiation-enhanced accumulation. () Immunohistochemistry of MF7GFP-IBD xenograft tumors depicting the decrease in endothelium (D31, brown), pericytes (α-sm, brown), and basal lamina (ollagen IV, brown) following imatinib mesylate treatment. Purple = hematoxylin, nuclei. Scale bar 5
6 = 200 µm. (B) Relative quantification of IH staining in MF7 GFP-IBD tumor sections. % area denotes the area of an image stained calculated using an ImageJ macro (details in methods). p 0.05 relative to control, p 0.05 relative to only, n = 3. () MF7 GFP-IBD tumor retention of ngiosense, measured using IVIS fluorescence quantification, is increased following treatment with imatinib mesylate alone, but not in conjunction with IR. ngiosense was administered 3 days after IR. p 0.05 relative to control, p 0.05 relative to only, ^ p 0.05 relative to imatinib mesylate only, at the same time point, n = 5. 6
7 No-IR No-IR IR IR B 0.25 Radiant Efficiency (x 10 9 ) tumor liver spleen heart Supplementary Figure 4. Preferential retention of Doxil in irradiated areas of tumors compared to other tissues. () MF7 GFP-IBD tumors (prior to Doxil injection, left panel), half irradiated in order to use the unirradiated half as a control, were administered Doxil (10mg/kg) with radiation-enhanced delivery leading to greater drug distribution in the 15 Gy-treated half (right panel). (B) Tissue retention of Doxil 26 days after i.v. administration in BLB/c female mice with TUBO hindlimb tumors. Doxil (10 mg/kg) was administered three days following tumor irradiation. n = 2 for each data set. 7
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Figure S1. B % of Phosphorylation 32H. 32ss
Figure S1 8H 32ss 32H 32Hc % of Phosphorylation 3 32H 2 1 32ss 1 2 3 4 Extract (μg) C % of Phosphorylation 18 12 6-32H 32Hc 8H 32ss Dbait Figure S1. List of the Dbait molecules and activation of DN-PK
Supplementary Information. Cofilin Regulates Nuclear Architecture through a Myosin-II Dependent Mechanotransduction Module
Supplementary Information Cofilin Regulates Nuclear Architecture through a Myosin-II Dependent Mechanotransduction Module O Neil Wiggan, Bryce Schroder, Diego Krapf, James R. Bamurg and Jennifer G. DeLuca
Supplementary Figure S1 Supplementary Figure S2
Supplementary Figure S A) The blots shown in Figure B were qualified by using Gel-Pro analyzer software (Rockville, MD, USA). The ratio of LC3II/LC3I to actin was then calculated. The data are represented
Nature Immunology: doi: /ni.3412
Supplementary Figure 1 Gata1 expression in heamatopoietic stem and progenitor populations. (a) Unsupervised clustering according to 100 top variable genes across single pre-gm cells. The two main cell
Mitosis. Single Nano Micro Milli Macro. Primary. PCNA expression
a b c DAPI YFP CC3 DAPI YFP PCNA DAPI YFP ph3 DAPI YFP KI67 e 6 Mitosis f 1 PCNA expression %ph3 + /YFP + n= 63 87 61 3 13 8 n= 15 3 9 1 5 %PCNA+/YFP+ 8 6 Supplementary Figure 1. Proliferation/apoptosis