JFIFWWCreated with GIMP~Photoshop 3.08BIMxCreated with GIMP8BIM%&R&[c՗=2W ^#8BIMW[#W[#8BIM&?8BIM 8BIM8BIM 8BIM 8BIM' 8BIMH/fflff/ff2Z5-8BIMp8BIM@@8BIM8BIMW v >pone.0106601.g004 > vnullboundsObjcRct1Top longLeftlongBtomlong vRghtlong >slicesVlLsObjcslicesliceIDlonggroupIDlongoriginenum ESliceOrigin autoGeneratedTypeenum ESliceTypeImg boundsObjcRct1Top longLeftlongBtomlong vRghtlong >urlTEXTnullTEXTMsgeTEXTaltTagTEXTcellTextIsHTMLboolcellTextTEXT horzAlignenumESliceHorzAligndefault vertAlignenumESliceVertAligndefault bgColorTypeenumESliceBGColorTypeNone topOutsetlong leftOutsetlong bottomOutsetlong rightOutsetlong8BIM( ?8BIM8BIM8BIM T8JFIFHH Adobe_CMAdobed            " ?   3!1AQa"q2B#$Rb34rC%Scs5&DTdE£t6UeuF'Vfv7GWgw5!1AQaq"2B#R3$brCScs4%&5DTdEU6teuFVfv'7GWgw ?ӝ2ɷ+:v:ϧU;Cvc7_}]IR:Vך]CRZk7JnI)I$JRK?gdWv9Mm s/slAÁw;7.[_7N7Ԕ}Yg}?}+ eX$fd̜Wc?өksѤI([mt}ى>%%3IU=K<3| Fk?wGnݸ%C+:uUX=f;Z]ll` 89/c aY)iI%?Uf![nOp,߻M fDVU7lmsٵ>^u0g qYv]>bJyoΒ3zS.}B7n;rݔ_Wn޻{N~쌑AotПSe$IJI$R{jί"ɨ6L;ճwsモ) z naoh nu$7:[ڜg蠟=Y378Vyysl;~v+T._V{伱Fs˽K*{RS-sM\:Iϻu7ߕmipSe;gl8'y H@IO>ߴ5q37;{~n#I$_=G~n6uwӼnwcU#?*9#3&Vqz vw~~Ĕ{NΧԲ?m4KO?3,=[щ;c{)) zō.&!Vќ汥!h8nm_gS}.׻n,sW:5U=k\Ymktn1%=2Ht:TKŕӗnDIM\Wdhoek_w?l@Sȋ-u{cԮo~T\×X|PZ7nu,kX1>QS?LJj>QS?LJjJ}&QG֦ ̗sr4nop^Wusky<ۼy?(_Z))b]Sd4{8io)I$JU^Ammn6[vvemqVZ؇~Xc;'MJy΅rl=?Y{-fKs#̷ҺmmK}=?M77Y.lf^is\g2ʚ?B? I%u'k#~I:]8ȑ2H^:r9.S?2C_oV@~jup+8G Gϥ}D?Zּ2n-Um'e7Z{9[Wo.gm RI$cYsNZk@_m(5@zMnL~ܼVV:`.k_n{Ek j&DSwWt{IO}@C}eٝMOdAu5;ۧ=qP?ct_?|o}g;}X]@61Aco%oQ8>n,7CMv j?IOc0UuMsk{.6oX\ү__XIJ^K6:Z?Yq9t :js-vn\K?YDz:QXUk}/gUԏOgEYU0-沶dX9ojX7bmҊ@zνk>vEisK@s՟yZF7K˺/qQcǵ\Չ3>w[]>~԰*ɯ:&ְS 롯ٱ˞Y?7b-ҊP Q.~}`Ӳ:nV6=~cf{C[ڽxw}o/N+edP_}Vcb{kJI$SUJrcpϩ`cTlg4SQe`cƻ힕ݻ*ӵgWA?V.;&p(gdО^ᕖߺ79mk~fgfͬ{\X77%

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Cells were lysed and viability was assessed by direct microscopy and counting intact nuclei. (B) WT and PON2 def cortical neurons were transfected with plasmid expressing Myc-PON2 and GFP (under independent promoters), or GFP as control, and subjected to 20 µM MPP+ for 48 hours. Cells were fixed and the nuclei were stained with Hoechst. Survival percentage represents the ratio of GFP-expressing cells with morphologically intact nuclei (D, a and b) to the total number of GFP positive cells. (C) WT and DJ-1 KO cortical neurons over-expressing PON2 and GFP or GFP alone as control (using adenovirus expressing PON2 or GFP) were subjected to 20 µM MPP+ for 48 hours. The survival assay was performed as described in part B. (D) Representative image of GFP positive neurons (a and c), and Hoechst-stained surviving (b) and dead (d) nuclei. (E) Western blot analysis of PON2 levels in PON2 deficient (PON2 def) and WT MEFs and also in WT MEFs infected with PON2-expressing adenovirus (WT+PON2 AV). The membrane was probed with PON2 antibody. (F) Western blot analysis for Myc in WT MEFs expressing control (Ctr) or Myc-PON2 plasmids. The Western blot was analyzed by Myc antibody. Statistical significance was assessed by Anova and post-hoc test Tukey on data obtained from three independent experiments (n = 3). * denotes p<0.05, **denotes p<0.01 and *** denotes p<0.001. info:doi/info:doi/10.1371/journal.pone.0106601.g004 Public Library of Science Figure 4 Creative Commons Attribution License info:doi/10.1371/journal.pone.0106601 C       C "  l   !18 "AQv7VXau#26qtw$3BWr5Rs(Cbf%&4FTdDESUce2!21QAaq"3B# ?ژץ@5 F2^vׅ[KO%$Wқ \l{dst ŖLWԪ=u ;ғ6闑 BϸtAT[NM>t]Y0ltE,H#7K5.ܨۭڥ=-UFT3jJvH)%~1#ΰsfu➗c[ejLBL_wߘǗΰ82w"*XL*ɜN6[lJ"Sf3-ű(g"H>c˃]9=8_~nEÿ^cՅM` j,HQʡp·CaDj6Rq$ϒylhT[ymOj{->E2j-ąf\O.BCE79t*nG9|Iji8q\\\['}7&dX޸΋`cbL 9jq)Y4Sg-(RO7׹lE_wIYy6Ji3EOJkr?|qOЎ2ƓX-oV,Z`aʐ5!]Bki1ZںBMlhP:9_6}Qj;^[ u.!}e~= ҏu7,[lK veSjq}”-$Fd|8tm?lKzdJ '&=+gwHz_n 4j-.'Lm;J3ȿ um|{'(\[hWmv uFSn2mx&mHվڙZjzLu(u'bRC*2ϙ^""3d"iu[pmTLTވRR!f\&6#ܔD|d #T[T;*j) mnG¤7DfۯE>g ÄuS5 1q\YDWetrl8NI6ֲYhJ^ x'`ù@G-}&❉:2y]bbjBȔ[IQn\ˑ)n)n*#{w(X=PjmTZȐޑjW E{$a 7 l؂jx>*"a'mJJ(C(-Z½2EDoWXڛSn⪾3%2VE:DK"Gr0jt<'jUzh1.'dIu-oRs.hHP03$H^mFG)&d{.pTSpTSpTSpTSpTSpTSpTSpTSpTSpTSpTSpTSpTSpTSpTko]]\Ӧl/X6gMS!5+o!$EȈ#7ck+.d-/cDT[QGBX3]*'J/!:ֳ&̄TrJUQ}K.."p\'Ḇ[[xZ>JX.0CK)327fχnB^bU)J|[Jx5.w@;䤉ni%ۆ(b>ku:WqD_=oҾs$izruM\!*RM 6DfF\*[_?iz+'cUؤRTJ3'\HH\-VCzK~Ӓ. 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