Author Manuscript Published OnlineFirst on September 11, 2015; DOI: 10.1158/0008-5472.CAN-14-3636 Author manuscripts have been peer reviewed and accepted for publication but have not yet been edited. Elevated expression of the C-type lectin CD93 in the glioblastoma vasculature regulates cytoskeletal rearrangements that enhance vessel function and reduce host survival *'1#,%#,)+.SQ#'&,%SQWQ- #023%,-SQWQ33,%SQ+"-0*1'0 3*&11,TQ 0' #-0%,)'SQ!#1+"880SQ$-&,$SQ #-0%#0#,"#*#, 30%UQ%,3111,"SQ (0#"0'))-,2M,SQ,(,+'21TQVQ,,.'+ #0%S# 1. Department of Immunology, Genetics and Pathology, The Rudbeck Laboratory, Uppsala University, Uppsala, Sweden 2. Department of Neuroscience, Neurology, University Hospital Uppsala, Uppsala, Sweden 3. Dept. of Neurology, University Medicine Goettingen, Robert-Koch Str. 40, 37075 Göttingen, Germany 4. Department of Neurology, Danish Epilepsy Center, Dianalund, Denmark 5. These authors contributed equally to the work. #0-00#1.-,"',%32&-0S,,.'+ #0%Q.#.02+#,2-$2++3,-*-%7Q #,#2'!1,")2&-*-%7Q 3" #!) -02-07Q5..1*5,'4#01'27Q,VYWSZW5..1*Q,5#"#,T)&-,#S&VXYRTSXXV[X (6S&VXSZWWZ[US+'*S,,T.'+ #0%'%.T33T1# Running title 0.[U0#%3*2#1,%'-%#,#1'1',%*'- *12-+ Key words %*'- *12-+Q,%'-%#,#1'1Q0V27.#*#!2',Q#,"-2&#*'*!#**"&#1'-,Q41!3*0$3,!2'-, Conflicts of interest &#32&-01&4#,-!-,$*'!2-$',2#0#122-"'1!*-1#T 1 Downloaded from cancerres.aacrjournals.org on October 1, 2021. © 2015 American Association for Cancer Research. Author Manuscript Published OnlineFirst on September 11, 2015; DOI: 10.1158/0008-5472.CAN-14-3636 Author manuscripts have been peer reviewed and accepted for publication but have not yet been edited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ownloaded from cancerres.aacrjournals.org on October 1, 2021. © 2015 American Association for Cancer Research. Author Manuscript Published OnlineFirst on September 11, 2015; DOI: 10.1158/0008-5472.CAN-14-3636 Author manuscripts have been peer reviewed and accepted for publication but have not yet been edited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ownloaded from cancerres.aacrjournals.org on October 1, 2021. © 2015 American Association for Cancer Research. Author Manuscript Published OnlineFirst on September 11, 2015; DOI: 10.1158/0008-5472.CAN-14-3636 Author manuscripts have been peer reviewed and accepted for publication but have not yet been edited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in vitroQ"3#2-"#$#!21',!72-1)#*#2*0#V00,%#+#,2,"*-11-$#,"-2&#*'*"&#0#,!#(3,!2'-,1T 2+.-02,2*7Q&'%&0.[U#6.0#11'-,',23+-04#11#*11'%,'$'!,2*7!-00#*2#"5'2&1&-02#0 1304'4*',!*','!*!-&-02-$.2'#,215'2&%0"#222V2D120-!72'!%*'-+1T*2-%#2&#0Q-30"2 '+.*72&20.[U'1)#70#%3*2-0-$%*'-+,%'-%#,#1'1,"41!3*0$3,!2'-,2&0-3%&2&# !-,20-*-$!#**V!#**,"!#**V+20'6"&#1'-,T 4 Downloaded from cancerres.aacrjournals.org on October 1, 2021. © 2015 American Association for Cancer Research. Author Manuscript Published OnlineFirst on September 11, 2015; DOI: 10.1158/0008-5472.CAN-14-3636 Author manuscripts have been peer reviewed and accepted for publication but have not yet been edited. MATERIALS & METHODS Tissue Microarray and Image analysis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umor cell and primary endothelial cell and pericytes culture TVS$' 0-10!-+Q"SXT(SR+#*,-++#0'!,7.#03*230#0-**#!2'-,Q ,111QDQ5,4' 0,2,"0"1Q," TXS%*'-+!#**1)',"%'$2$0-+.0T #8,$0,7Q C2Q"3".#12#,%',##0#"2-#6.0#11*3!'$#01#',&-31#&,%et alTQ3,.3 *'1&#"5#0# !3*230#"-,SRV!+!3*230#"'1&#1',.3* #!!-_1-"'$'#"%*##"'3+'$##!&,-*-%'#1Q 00*1 "Q0Q5,13..*#+#,2#"5'2&SR$$#2*!*$1#03+(0,R,'%+V*"0'!&Q,2T-3'1QFQ 5,2UY-0,"W$0FT [W$'0',&3+'"'$'#"!&+ #0T 3+,.#0+*'!0-41!3*0,"-2&#*'*0#**1.0RU"'-+#"'!*Q5..1*Q ,5#"#,Q,")0-+-0#**Q#'"#* #0%Q #0+,75#0#!3*230#"3.2-.11%#SW-,%#*2',V!-2#" !3*230#"'1&#1',,"-2&#*'*0#**"1*#"'3+5'2&$3**13..*#+#,21"VDTQ)0-+-0#**2 UY-0,"W$0FT [W$'0',&3+'"'$'#"!&+ #0T3+,"0',)#0'!72#1")RU 5 Downloaded from cancerres.aacrjournals.org on October 1, 2021.