Journal: Journal of Neuroscience

Article Title: RAS/ERK Signaling Controls Proneural Genetic Programs in Cortical Development and Gliomagenesis

doi: 10.1523/jneurosci.4077-13.2014

Figure Lengend Snippet: Figure4. RAS/ERKsignalinginhibitscorticalneurogenesis.A–L,E12.53E15.5electroporationsofapCIG2controlvector(A,A,E,E,I,I),RasV12(B,B,F,F,J,J),bRafV600E(C,C,G,G, K,K),andMekCA(D,D,H,H,L,L)vectors(expressingGFP).TransfectedbrainswereanalyzedforcoexpressionofGFP(green)withpan-neuronalmarkerNeuN(red;A–D,A–D)andthecortical neuronal marker Tbr1 (red; E–H, E–H), or for transcripts for GFP (I–L) or the subcortical neuronal marker Dlx1 (I–L). Dashed lines outline the transfected region in the neocortex. Yellow arrowheads mark transfected areas with reduced expression of NeuN (B, C, D), and Tbr1 (F, G, H), and red arrowheads mark ectopic expression of Dlx1 (L). Scale bars: 250 m. M, N, QuantificationofGFP cellscoexpressingNeuN(M)orTbr1(N).pvaluesofpCIG2-RasV12,pEF-RasV12,bRafV600E,MekCA,AktCA,RalAV23,andRalBQ72EarerelativetopCIG2control,whereasp values of pCIG2RasV12pCIG2MekDN, pCIG2RasV12pCIG2AktDN, pCIG2Rasv12pCIG2RalADN, and pCIG2RasV12pCIG2RalBDN are relative to pCIG2RasV12. *p 0.05, **p 0.01, ***p 0.005. ctx, neocortex; str, striatum.

Article Snippet: Huang, Addgene plasmid 15269; B. P. Zhou et al., 2000), RalAV23 (from W. Hahn, Addgene plasmid 15252; Sablina et al., 2007), RalADN (RalAS31N, from C. Counter, Addgene plasmids 19718; Lim et al., 2005), RalBQ72L (from C. Counter, Addgene plasmid 19721; Lim et al., 2005), and RalBDN (RalBS28N, from C. Counter, Addgene plasmid 19722; Lim et al., 2005) were obtained from Addgene.

Techniques: Marker, Transfection, Expressing

Journal: Journal of Neuroscience

Article Title: RAS/ERK Signaling Controls Proneural Genetic Programs in Cortical Development and Gliomagenesis

doi: 10.1523/jneurosci.4077-13.2014

Figure Lengend Snippet: Figure 6. Activation of RAS signaling in cortical progenitors results in tumorigenesis. A–E, E18.5 control (C) and E12.53E18.5 brains electroporated with pCIG2RasV12 (A, B, D, E, E) were processed for H&E staining (B–E, E), or GFP transcripts (A). F–H, E12.53E16.5 electroporations of a pCIG2 control vector (F–F) and pCIG2RasV12 (G–G, H–H). Transfected brains were analyzed for coexpression of GFP (green), proliferation marker Ki67 (red), and pan-neuronal marker Tuj1 (blue). White arrowheads mark RasV12 transfected cells expressing Ki67 instead of Tuj1 (H–H).I–P,E12.53E15.5brainselectroporatedwithapCIG2controlvector(I,I,K,K,M,M,O,O)orpCIG2RasV12(J,J,L,L,N,N,P,P)wereprocessedforimmunostainingofpan-neural progenitormarkersSox2(I,I,J,J)andNestin(K,K,L,L),cortical-specificprogenitormarkersPax6(M,M,N,N),andTbr2(O,O,P,P).RedarrowheadsmarkectopicexpressionofSox2(J) and Nestin (L), while yellow arrowheads mark transfected areas with reduced expression of Pax6 (N) and Tbr2 (P). Q–V, E12.53E15.5 electroporations of pCIG2RasV12 with (T–V, V) or withoutNeurog2(Q–S,S).TransfectedbrainswereanalyzedforthetranscriptsofSpry2(Q,T),Pdgfa(R,U),orcoexpressionofGFPandTuj1(S,S,V,V).Redarrowheadsmarkectopicexpression ofSpry2(Q,T)orPdgfa(R).W–Z,E12.53E15.5electroporationsofpCIG2(W,W),AktA(X,X),RalAV23(Y,Y),andRalBQ72L(Z,Z)followedbyanalysisofBrdUincorporation.Redarrowheads markectopicBrdUincorporation.Dashedlinesoutlinethetransfectedregionintheneocortex.Scalebars:I–V,I–V,F–F,G–G250m;H–H,67.5m.CP,corticalplate;ctx,neocortex;str,striatum.

Article Snippet: Huang, Addgene plasmid 15269; B. P. Zhou et al., 2000), RalAV23 (from W. Hahn, Addgene plasmid 15252; Sablina et al., 2007), RalADN (RalAS31N, from C. Counter, Addgene plasmids 19718; Lim et al., 2005), RalBQ72L (from C. Counter, Addgene plasmid 19721; Lim et al., 2005), and RalBDN (RalBS28N, from C. Counter, Addgene plasmid 19722; Lim et al., 2005) were obtained from Addgene.

Techniques: Activation Assay, Control, Staining, Plasmid Preparation, Transfection, Marker, Expressing