U19 Hope Lab June 2015
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Transcript of U19 Hope Lab June 2015
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U19 conference call
Thomas Hope labProject progress update
Katarina Kotnik Halavaty
June 1! "1#
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$esults of the "nd group ofanimals
•
% u c i f e r a s e
a c
t i v i t y
•
& i r s t t r a n s d u c e d
c e l l s
•
P r o v i r a l ' ( )
d e t e c
t i o n
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Uterusovaries
*ervi+
,agina
%o-er ,agina
Upper ,agina
)nalyses.
/nested P*/0uorescenmicroscopy
&reeing of tissue
2hole &$T
• Tissue is soaked in luciferin for 1 minutes• 'etection of luciferase activity in &$T using an 3n ,ivo 3maging 4ystem
53,346
ssue uc erase act v tyscreen
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T'& 3,$ 7 7 7 7 7 7 /
)nimals 889: 88#:# 88#;< 881:;< 88#"<
?vary - + ++ - - - -
Uterus - - - - - - -
*ervi+ - - - - - - -
,agina - - - - - - -
4ummary of luciferase activity
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m*herry %uciferase@*A# Berge
m*herry *';@&3T* Berge
Transduced cell in an ovary of acontrol pigtail macaCue
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m*herry@%uciferase merge
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T'& 3,$ 7 7 7 7 7 7 /
)nimals 889: 88#:# 88#;< 881:;< 88#"<
?vary 1 9 1 0 3
?vary "
4ummary of transduced cells inovaries
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(o %T$ nested P*$ for PigtailBacaCues
1st P*$ product. 1># kb"nd P*$ product. "# bp
mCherry primers
1st P*$ product. # bp"nd P*$ product. ;# bp
LTR primers LTR primers
*B,%T$%uciferas
e3$D4 %T$m*herry
2P$D
These animals -ere already challenged -ith 4H3,1"p; live virusE
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1>
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T'& 3,$ 7 7 7 7 7 7 /
)nimals 889: 88#:# 88#;< 881:;< 88#"<
?vary 1 - - - + - - +
?vary " - - - -
&allopian tubes
- + - -
*ervi+ - - + - + - -
,agina - - - - - - -
4ummary of proviral '() detection
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*onclusions
• %uciferase activity -as detected in. ovaries of t-o T'& 3,$ treated pigtail macaCues
• 4ingle infected cells -ere found in.
ovaries of t-o T'& 3,$ treated pigtail macaCues an ovary of a control animal
• Proviral '() -as detected by nested P*$ in.
an ovary of T'& 3,$ treated animal an ovary of a control animal
*ervical tissue of t-o T'& 3,$ treated animals
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'ata of the :rd group ofanimals
% u
c
i f e r
a s e
a c t i
v i t y
Animal code88:19
88;"
88:1"
1>
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Preluciferin Postluciferin
!!319 "hole FRTPlacebo ring
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Postluciferin
!!319 #$aries
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Preluciferin Postluciferin
!!0%&' "hole FRT
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Postluciferin
!!0%&' #$aries
?variescut in half
andreimaged
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Preluciferin Postluciferin
!!031' "hole FRT
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Postluciferin
!!031' #$aries
?varies cut in half and reimaged
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&allopian tubes
?varian fat
!!031'&allopian Tubes and ovarian fat
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Preluciferin Postluciferin
1(&))* "hole FRT
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1(&))* #$aries and Fallopian ,es
?varies&allopian tubes
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%uciferase activity summary
IVR + + + +
TDF - + + +
Animalcode
!!319 !!0%&' !!031' 1(&))*
#$aries + ++ + ++
.er,s - - - -
Cer$i/ - - - -
Vagina - - - -
Laia - - - -