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mso-tstyle-shading:white; mso-tstyle-shading-themecolor:background1; mso-tstyle-border-right:.5pt solid #7F7F7F; mso-tstyle-border-right-themecolor:text1; mso-tstyle-border-right-themetint:128; text-align:right; font-size:13.0pt; mso-ansi-font-size:13.0pt; font-family:"Calibri Light",sans-serif; mso-ascii-font-family:"Calibri Light"; mso-ascii-theme-font:major-latin; mso-fareast-font-family:"Times New Roman"; mso-fareast-theme-font:major-fareast; mso-hansi-font-family:"Calibri Light"; mso-hansi-theme-font:major-latin; mso-bidi-font-family:"Times New Roman"; mso-bidi-theme-font:major-bidi; mso-ansi-font-style:italic; mso-bidi-font-style:italic;} table.MsoTable15Plain5LastCol {mso-style-name:"Plain Table 5"; mso-table-condition:last-column; mso-style-priority:45; mso-style-unhide:no; mso-tstyle-shading:white; mso-tstyle-shading-themecolor:background1; mso-tstyle-border-left:.5pt solid #7F7F7F; mso-tstyle-border-left-themecolor:text1; mso-tstyle-border-left-themetint:128; font-size:13.0pt; mso-ansi-font-size:13.0pt; font-family:"Calibri Light",sans-serif; mso-ascii-font-family:"Calibri Light"; mso-ascii-theme-font:major-latin; mso-fareast-font-family:"Times New Roman"; mso-fareast-theme-font:major-fareast; mso-hansi-font-family:"Calibri Light"; mso-hansi-theme-font:major-latin; mso-bidi-font-family:"Times New Roman"; mso-bidi-theme-font:major-bidi; mso-ansi-font-style:italic; mso-bidi-font-style:italic;} table.MsoTable15Plain5OddColumn {mso-style-name:"Plain Table 5"; mso-table-condition:odd-column; mso-style-priority:45; mso-style-unhide:no; mso-tstyle-shading:#F2F2F2; mso-tstyle-shading-themecolor:background1; mso-tstyle-shading-themeshade:242;} table.MsoTable15Plain5OddRow {mso-style-name:"Plain Table 5"; mso-table-condition:odd-row; mso-style-priority:45; mso-style-unhide:no; mso-tstyle-shading:#F2F2F2; mso-tstyle-shading-themecolor:background1; mso-tstyle-shading-themeshade:242;} table.MsoTable15Plain5NECell {mso-style-name:"Plain Table 5"; mso-table-condition:ne-cell; mso-style-priority:45; mso-style-unhide:no; mso-tstyle-border-left:cell-none;} table.MsoTable15Plain5NWCell {mso-style-name:"Plain Table 5"; mso-table-condition:nw-cell; mso-style-priority:45; mso-style-unhide:no; mso-tstyle-border-right:cell-none;} table.MsoTable15Plain5SECell {mso-style-name:"Plain Table 5"; mso-table-condition:se-cell; mso-style-priority:45; mso-style-unhide:no; mso-tstyle-border-left:cell-none;} table.MsoTable15Plain5SWCell {mso-style-name:"Plain Table 5"; mso-table-condition:sw-cell; mso-style-priority:45; mso-style-unhide:no; mso-tstyle-border-right:cell-none;} </style><![endif]--><!--[if gte mso 9]><xml><o:shapedefaults v:ext="edit" spidmax="1026"/></xml><![endif]--><!--[if gte mso 9]><xml><o:shapelayout v:ext="edit"><o:idmap v:ext="edit" data="1"/></o:shapelayout></xml><![endif]--></head><body lang=EN-US link=blue vlink="#954F72" style='tab-interval:.5in'><div class=WordSection1><p class=MsoNormal>Meeting with Kota Kaneko 3/17/2016<p class=MsoNormal>Kota performed a FACS experiment on Wednesday 3/16/2016 on a single mouse.<p class=MsoListParagraphCxSpFirst style='text-indent:-.25in;mso-list:l0 level1 lfo3'><![if !supportLists]><span style='font-family:Symbol;mso-fareast-font-family:Symbol;mso-bidi-font-family: Symbol'><span style='mso-list:Ignore'>路<span style='font:7.0pt "Times New Roman"'>聽聽聽聽聽聽聽聽 </span></span></span><![endif]>Perfused liver with PBS to flush RBCs and platelets via portal vein<p class=MsoListParagraphCxSpMiddle style='text-indent:-.25in;mso-list:l0 level1 lfo3'><![if !supportLists]><span style='font-family:Symbol;mso-fareast-font-family:Symbol;mso-bidi-font-family: Symbol'><span style='mso-list:Ignore'>路<span style='font:7.0pt "Times New Roman"'>聽聽聽聽聽聽聽聽 </span></span></span><![endif]>Perfused with collagenase I and IV and dissociated with razor<p class=MsoListParagraphCxSpMiddle style='text-indent:-.25in;mso-list:l0 level1 lfo3'><![if !supportLists]><span style='font-family:Symbol;mso-fareast-font-family:Symbol;mso-bidi-font-family: Symbol'><span style='mso-list:Ignore'>路<span style='font:7.0pt "Times New Roman"'>聽聽聽聽聽聽聽聽 </span></span></span><![endif]>Spun down hepatocytes at 50 g for 5 min<p class=MsoListParagraphCxSpMiddle style='text-indent:-.25in;mso-list:l0 level1 lfo3'><![if !supportLists]><span style='font-family:Symbol;mso-fareast-font-family:Symbol;mso-bidi-font-family: Symbol'><span style='mso-list:Ignore'>路<span style='font:7.0pt "Times New Roman"'>聽聽聽聽聽聽聽聽 </span></span></span><![endif]>Stained with Hoerst and PI with Verapamil (to block calcium channel, prevent pumping out of Hoerst)<p class=MsoListParagraphCxSpMiddle style='margin-left:1.0in;mso-add-space: auto;text-indent:-.25in;mso-list:l0 level2 lfo3'><![if !supportLists]><span style='font-family:"Courier New";mso-fareast-font-family:"Courier New"'><span style='mso-list:Ignore'>o<span style='font:7.0pt "Times New Roman"'>聽聽 </span></span></span><![endif]>Live cells stain with Hoerst but not PI<p class=MsoListParagraphCxSpMiddle style='margin-left:1.0in;mso-add-space: auto;text-indent:-.25in;mso-list:l0 level2 lfo3'><![if !supportLists]><span style='font-family:"Courier New";mso-fareast-font-family:"Courier New"'><span style='mso-list:Ignore'>o<span style='font:7.0pt "Times New Roman"'>聽聽 </span></span></span><![endif]>Dead cells stain for both<p class=MsoListParagraphCxSpLast style='text-indent:-.25in;mso-list:l0 level1 lfo3'><![if !supportLists]><span style='font-family:Symbol;mso-fareast-font-family:Symbol;mso-bidi-font-family: Symbol'><span style='mso-list:Ignore'>路<span style='font:7.0pt "Times New Roman"'>聽聽聽聽聽聽聽聽 </span></span></span><![endif]>Sorted at Sanford HESSCF on FACScanto:<p class=MsoNormal><o:p>聽</o:p><p class=MsoNormal>The Feng group has a variety of livor tumor models:<p class=MsoListParagraphCxSpFirst style='text-indent:-.25in;mso-list:l0 level1 lfo3'><![if !supportLists]><span style='font-family:Symbol;mso-fareast-font-family:Symbol;mso-bidi-font-family: Symbol'><span style='mso-list:Ignore'>路<span style='font:7.0pt "Times New Roman"'>聽聽聽聽聽聽聽聽 </span></span></span><![endif]>Albumin-Cre mice (B6.Cg-Tg(Alb-cre)21Mgn/J) are crossed with a variety of floxed strains<p class=MsoListParagraphCxSpMiddle style='margin-left:1.0in;mso-add-space: auto;text-indent:-.25in;mso-list:l0 level2 lfo3'><![if !supportLists]><span style='font-family:"Courier New";mso-fareast-font-family:"Courier New"'><span style='mso-list:Ignore'>o<span style='font:7.0pt "Times New Roman"'>聽聽 </span></span></span><![endif]>Crossed with flox-stop GFP reporter to validate specificity of knockout<p class=MsoListParagraphCxSpMiddle style='margin-left:1.0in;mso-add-space: auto;text-indent:-.25in;mso-list:l0 level2 lfo3'><![if !supportLists]><span style='font-family:"Courier New";mso-fareast-font-family:"Courier New"'><span style='mso-list:Ignore'>o<span style='font:7.0pt "Times New Roman"'>聽聽 </span></span></span><![endif]>Saw GFP in hepatocytes only<p class=MsoListParagraphCxSpMiddle style='text-indent:-.25in;mso-list:l0 level1 lfo3'><![if !supportLists]><span style='font-family:Symbol;mso-fareast-font-family:Symbol;mso-bidi-font-family: Symbol'><span style='mso-list:Ignore'>路<span style='font:7.0pt "Times New Roman"'>聽聽聽聽聽聽聽聽 </span></span></span><![endif]>Floxed PTEN and floxed Ptpn11 (SHP2), ~2 kb deletion of the promoter<p class=MsoListParagraphCxSpMiddle style='margin-left:1.0in;mso-add-space: auto;text-indent:-.25in;mso-list:l0 level2 lfo3'><![if !supportLists]><span style='font-family:"Courier New";mso-fareast-font-family:"Courier New"'><span style='mso-list:Ignore'>o<span style='font:7.0pt "Times New Roman"'>聽聽 </span></span></span><![endif]>Mouse Pten is on <a href="http://www.ensembl.org/Mus_musculus/Location/View?db=core;g=ENSMUSG00000013663;r=19:32757497-32826160"><span style='mso-bidi-font-family:Helvetica;color:windowtext;background:white; text-decoration:none;text-underline:none'>Chromosome 19: 32,757,497-32,826,160</span></a><span class=apple-converted-space><span style='mso-bidi-font-family:Helvetica; background:white'>聽</span></span><span style='mso-bidi-font-family:Helvetica; background:white'>forward strand.</span><p class=MsoListParagraphCxSpMiddle style='margin-left:1.0in;mso-add-space: auto;text-indent:-.25in;mso-list:l0 level2 lfo3'><![if !supportLists]><span style='font-family:"Courier New";mso-fareast-font-family:"Courier New"'><span style='mso-list:Ignore'>o<span style='font:7.0pt "Times New Roman"'>聽聽 </span></span></span><![endif]>Mouse Ptpn11 is on <a href="http://www.ensembl.org/Mus_musculus/Location/View?db=core;g=ENSMUSG00000043733;r=5:121130533-121191397"><span style='mso-bidi-font-family:Helvetica;color:windowtext;background:white; text-decoration:none;text-underline:none'>Chromosome 5: 121,130,533-121,191,397</span></a><span class=apple-converted-space><span style='mso-bidi-font-family:Helvetica; background:white'>聽</span></span><span style='mso-bidi-font-family:Helvetica; background:white'>reverse strand</span><p class=MsoListParagraphCxSpLast style='text-indent:-.25in;mso-list:l0 level1 lfo3'><![if !supportLists]><span style='font-family:Symbol;mso-fareast-font-family:Symbol;mso-bidi-font-family: Symbol'><span style='mso-list:Ignore'>路<span style='font:7.0pt "Times New Roman"'>聽聽聽聽聽聽聽聽 </span></span></span><![endif]><span style='mso-bidi-font-family:Helvetica; background:white'>DEN (Diethylnitrosamine) is also used to generate liver tumors</span><div align=center> {| class=MsoTable15Plain3 border=0 cellspacing=0 cellpadding=0 width=631 style='width:473.4pt;border-collapse:collapse;mso-yfti-tbllook:1184; mso-padding-alt:0in 5.4pt 0in 5.4pt'<tr style='mso-yfti-irow:-1;mso-yfti-firstrow:yes;mso-yfti-lastfirstrow:yes; height:12.75pt'><td width=67 valign=top style='width:50.3pt;border:none;border-bottom:solid #7F7F7F 1.0pt; mso-border-bottom-themecolor:text1;mso-border-bottom-themetint:128; mso-border-bottom-alt:solid #7F7F7F .5pt;mso-border-bottom-themecolor:text1; mso-border-bottom-themetint:128;padding:0in 5.4pt 0in 5.4pt;height:12.75pt'><p class=MsoNormal style='margin-bottom:0in;margin-bottom:.0001pt;line-height: normal;mso-yfti-cnfc:517'>'''<span style='text-transform:uppercase'>Model<o:p></o:p></span>'''<td width=134 valign=top style='width:100.65pt;border:none;border-bottom: solid #7F7F7F 1.0pt;mso-border-bottom-themecolor:text1;mso-border-bottom-themetint: 128;mso-border-bottom-alt:solid #7F7F7F .5pt;mso-border-bottom-themecolor: text1;mso-border-bottom-themetint:128;padding:0in 5.4pt 0in 5.4pt;height: 12.75pt'><p class=MsoNormal style='margin-bottom:0in;margin-bottom:.0001pt;line-height: normal;mso-yfti-cnfc:1'>'''<span style='text-transform:uppercase'>Condition 1<o:p></o:p></span>'''<td width=104 valign=top style='width:77.8pt;border:none;border-bottom:solid #7F7F7F 1.0pt; mso-border-bottom-themecolor:text1;mso-border-bottom-themetint:128; mso-border-bottom-alt:solid #7F7F7F .5pt;mso-border-bottom-themecolor:text1; mso-border-bottom-themetint:128;padding:0in 5.4pt 0in 5.4pt;height:12.75pt'><p class=MsoNormal style='margin-bottom:0in;margin-bottom:.0001pt;line-height: normal;mso-yfti-cnfc:1'>'''<span style='text-transform:uppercase'>Condition 2<o:p></o:p></span>'''<td width=326 valign=top style='width:244.65pt;border:none;border-bottom: solid #7F7F7F 1.0pt;mso-border-bottom-themecolor:text1;mso-border-bottom-themetint: 128;mso-border-bottom-alt:solid #7F7F7F .5pt;mso-border-bottom-themecolor: text1;mso-border-bottom-themetint:128;padding:0in 5.4pt 0in 5.4pt;height: 12.75pt'><p class=MsoNormal style='margin-bottom:0in;margin-bottom:.0001pt;line-height: normal;mso-yfti-cnfc:1'>'''<span style='text-transform:uppercase'>Result<o:p></o:p></span>''' |- style='mso-yfti-irow:0;height:25.5pt'<td width=67 valign=top style='width:50.3pt;border:none;border-right:solid #7F7F7F 1.0pt; mso-border-right-themecolor:text1;mso-border-right-themetint:128;mso-border-right-alt: solid #7F7F7F .5pt;mso-border-right-themecolor:text1;mso-border-right-themetint: 128;background:#F2F2F2;mso-background-themecolor:background1;mso-background-themeshade: 242;padding:0in 5.4pt 0in 5.4pt;height:25.5pt'><p class=MsoNormal style='margin-bottom:0in;margin-bottom:.0001pt;line-height: normal;mso-yfti-cnfc:68'>'''<span style='text-transform:uppercase'>A<o:p></o:p></span>'''<td width=134 valign=top style='width:100.65pt;background:#F2F2F2;mso-background-themecolor: background1;mso-background-themeshade:242;padding:0in 5.4pt 0in 5.4pt; height:25.5pt'><p class=MsoNormal style='margin-bottom:0in;margin-bottom:.0001pt;line-height: normal;mso-yfti-cnfc:64'>Pten KO<td width=104 valign=top style='width:77.8pt;background:#F2F2F2;mso-background-themecolor: background1;mso-background-themeshade:242;padding:0in 5.4pt 0in 5.4pt; height:25.5pt'><p class=MsoNormal style='margin-bottom:0in;margin-bottom:.0001pt;line-height: normal;mso-yfti-cnfc:64'>Shp2 KO<td width=326 valign=top style='width:244.65pt;background:#F2F2F2;mso-background-themecolor: background1;mso-background-themeshade:242;padding:0in 5.4pt 0in 5.4pt; height:25.5pt'><p class=MsoNormal style='margin-bottom:0in;margin-bottom:.0001pt;line-height: normal;mso-yfti-cnfc:64'>Fatty liver, tumorgenesis |- style='mso-yfti-irow:1;height:38.25pt'<td width=67 valign=top style='width:50.3pt;border:none;border-right:solid #7F7F7F 1.0pt; mso-border-right-themecolor:text1;mso-border-right-themetint:128;mso-border-right-alt: solid #7F7F7F .5pt;mso-border-right-themecolor:text1;mso-border-right-themetint: 128;padding:0in 5.4pt 0in 5.4pt;height:38.25pt'><p class=MsoNormal style='margin-bottom:0in;margin-bottom:.0001pt;line-height: normal;mso-yfti-cnfc:4'>'''<span style='text-transform:uppercase'>B<o:p></o:p></span>'''<td width=134 valign=top style='width:100.65pt;padding:0in 5.4pt 0in 5.4pt; height:38.25pt'><p class=MsoNormal style='margin-bottom:0in;margin-bottom:.0001pt;line-height: normal'>DEN<td width=104 valign=top style='width:77.8pt;padding:0in 5.4pt 0in 5.4pt; height:38.25pt'><p class=MsoNormal style='margin-bottom:0in;margin-bottom:.0001pt;line-height: normal'>Shp2 KO<td width=326 valign=top style='width:244.65pt;padding:0in 5.4pt 0in 5.4pt; height:38.25pt'><p class=MsoNormal style='margin-bottom:0in;margin-bottom:.0001pt;line-height: normal'>Hepatocyte death + inflamation, tumorgenesis |- style='mso-yfti-irow:2;height:12.75pt'<td width=67 valign=top style='width:50.3pt;border:none;border-right:solid #7F7F7F 1.0pt; mso-border-right-themecolor:text1;mso-border-right-themetint:128;mso-border-right-alt: solid #7F7F7F .5pt;mso-border-right-themecolor:text1;mso-border-right-themetint: 128;background:#F2F2F2;mso-background-themecolor:background1;mso-background-themeshade: 242;padding:0in 5.4pt 0in 5.4pt;height:12.75pt'><p class=MsoNormal style='margin-bottom:0in;margin-bottom:.0001pt;line-height: normal;mso-yfti-cnfc:68'>'''<span style='text-transform:uppercase'>C<o:p></o:p></span>'''<td width=134 valign=top style='width:100.65pt;background:#F2F2F2;mso-background-themecolor: background1;mso-background-themeshade:242;padding:0in 5.4pt 0in 5.4pt; height:12.75pt'><p class=MsoNormal style='margin-bottom:0in;margin-bottom:.0001pt;line-height: normal;mso-yfti-cnfc:64'>c-Met/beta-Cat OE<td width=104 valign=top style='width:77.8pt;background:#F2F2F2;mso-background-themecolor: background1;mso-background-themeshade:242;padding:0in 5.4pt 0in 5.4pt; height:12.75pt'><p class=MsoNormal style='margin-bottom:0in;margin-bottom:.0001pt;line-height: normal;mso-yfti-cnfc:64'>na<td width=326 valign=top style='width:244.65pt;background:#F2F2F2;mso-background-themecolor: background1;mso-background-themeshade:242;padding:0in 5.4pt 0in 5.4pt; height:12.75pt'><p class=MsoNormal style='margin-bottom:0in;margin-bottom:.0001pt;line-height: normal;mso-yfti-cnfc:64'>Tumorgenesis |- style='mso-yfti-irow:3;height:25.5pt'<td width=67 valign=top style='width:50.3pt;border:none;border-right:solid #7F7F7F 1.0pt; mso-border-right-themecolor:text1;mso-border-right-themetint:128;mso-border-right-alt: solid #7F7F7F .5pt;mso-border-right-themecolor:text1;mso-border-right-themetint: 128;padding:0in 5.4pt 0in 5.4pt;height:25.5pt'><p class=MsoNormal style='margin-bottom:0in;margin-bottom:.0001pt;line-height: normal;mso-yfti-cnfc:4'>'''<span style='text-transform:uppercase'>D<o:p></o:p></span>'''<td width=134 valign=top style='width:100.65pt;padding:0in 5.4pt 0in 5.4pt; height:25.5pt'><p class=MsoNormal style='margin-bottom:0in;margin-bottom:.0001pt;line-height: normal'>c-Met/beta-Cat OE<td width=104 valign=top style='width:77.8pt;padding:0in 5.4pt 0in 5.4pt; height:25.5pt'><p class=MsoNormal style='margin-bottom:0in;margin-bottom:.0001pt;line-height: normal'>Shp2 KO<td width=326 valign=top style='width:244.65pt;padding:0in 5.4pt 0in 5.4pt; height:25.5pt'><p class=MsoNormal style='margin-bottom:0in;margin-bottom:.0001pt;line-height: normal'>Hepatocyte death + inflamation |- style='mso-yfti-irow:4;mso-yfti-lastrow:yes;height:38.25pt'<td width=67 valign=top style='width:50.3pt;border:none;border-right:solid #7F7F7F 1.0pt; mso-border-right-themecolor:text1;mso-border-right-themetint:128;mso-border-right-alt: solid #7F7F7F .5pt;mso-border-right-themecolor:text1;mso-border-right-themetint: 128;background:#F2F2F2;mso-background-themecolor:background1;mso-background-themeshade: 242;padding:0in 5.4pt 0in 5.4pt;height:38.25pt'><p class=MsoNormal style='margin-bottom:0in;margin-bottom:.0001pt;line-height: normal;mso-yfti-cnfc:68'>'''<span style='text-transform:uppercase'>E<o:p></o:p></span>'''<td width=134 valign=top style='width:100.65pt;background:#F2F2F2;mso-background-themecolor: background1;mso-background-themeshade:242;padding:0in 5.4pt 0in 5.4pt; height:38.25pt'><p class=MsoNormal style='margin-bottom:0in;margin-bottom:.0001pt;line-height: normal;mso-yfti-cnfc:64'>Shp2 KO<td width=104 valign=top style='width:77.8pt;background:#F2F2F2;mso-background-themecolor: background1;mso-background-themeshade:242;padding:0in 5.4pt 0in 5.4pt; height:38.25pt'><p class=MsoNormal style='margin-bottom:0in;margin-bottom:.0001pt;line-height: normal;mso-yfti-cnfc:64'>na<td width=326 valign=top style='width:244.65pt;background:#F2F2F2;mso-background-themecolor: background1;mso-background-themeshade:242;padding:0in 5.4pt 0in 5.4pt; height:38.25pt'><p class=MsoNormal style='margin-bottom:0in;margin-bottom:.0001pt;line-height: normal;mso-yfti-cnfc:64'>Hepatocyte death + inflamation, some benign tumors |}</div><p class=MsoNormal><o:p>聽</o:p><p class=MsoNormal><o:p>聽</o:p></div></body></html>
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