- 34-3600 - Invitrogen Antibodies EFNB3 antibody

Submitted references A neuronal molecular switch through cell-cell contact that regulates quiescent neural stem cells.

Dong J, Pan YB, Wu XR, He LN, Liu XD, Feng DF, Xu TL, Sun S, Xu NJ
Science advances 2019 Feb;5(2):eaav4416

Retrograde regulation of mossy fiber axon targeting and terminal maturation via postsynaptic Lnx1.

Liu XD, Zhu XN, Halford MM, Xu TL, Henkemeyer M, Xu NJ
The Journal of cell biology 2018 Nov 5;217(11):4007-4024

Ephrin-B3 coordinates timed axon targeting and amygdala spinogenesis for innate fear behaviour.

Zhu XN, Liu XD, Sun S, Zhuang H, Yang JY, Henkemeyer M, Xu NJ
Nature communications 2016 Mar 24;7:11096

Anchoring and synaptic stability of PSD-95 is driven by ephrin-B3.

Hruska M, Henderson NT, Xia NL, Le Marchand SJ, Dalva MB
Nature neuroscience 2015 Nov;18(11):1594-605

EphA4-dependent axon retraction and midline localization of Ephrin-B3 are disrupted in the spinal cord of mice lacking mDia1 and mDia3 in combination.

Toyoda Y, Shinohara R, Thumkeo D, Kamijo H, Nishimaru H, Hioki H, Kaneko T, Ishizaki T, Furuyashiki T, Narumiya S
Genes to cells : devoted to molecular & cellular mechanisms 2013 Oct;18(10):873-85

Evaluation of Eph receptor and ephrin expression within the human cornea and limbus.

Hogerheyde TA, Stephenson SA, Harkin DG, Bray LJ, Madden PW, Woolf MI, Richardson NA
Experimental eye research 2013 Feb;107:110-20

Methylmercury alters Eph and ephrin expression during neuronal differentiation of P19 embryonal carcinoma cells.

Wilson DT, Polunas MA, Zhou R, Halladay AK, Lowndes HE, Reuhl KR
Neurotoxicology 2005 Aug;26(4):661-74

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 1 PSD-95 is in a complex with ephrin-B3 at synapses a) ST imulated E mission D epletion (STED) imaging of PSD-95 and ephrin-B3 at synapses. DIV21 tdTomato transfected rat cortical neurons were labeled with the indicated antibodies. PSD-95 and ephrin-B3 were imaged at super-resolution using STED (~80 nm). VGlut1 and tdTomato were imaged at conventional resolution (~250 nm) using confocal. Images are high contrast examples of PSD-95, ephrin-B3 and vGlut1 immunostaining. Arrows indicate PSD-95, ephrin-B3 and vGlut1 co-clusters in dendritic spines. Scale bar: 2 um. b) PSD-95, ephrin-B3 and vGlut1 are found in most (>=80%) spines (248 spines on 15 neurons, two separate experiments). c) PSD-95-ephrin-B3 co-clusters are found predominantly in vGlut + spines (162 PSD-95-ephrin-B3 containing spines out of 202 vGlut1 + spines). Single PSD-95 (14 out of 202 vGlut + spines) or ephrin-B3 (20 out of 202 vGlut + spines) clusters are rarely found at synapses. d-f) Co-immunoprecipitation (co-IP) of ephrin-B3 and PSD-95 from P21 wild type and Efnb3 -/- mouse whole brain lysates. The efficacy of ephrin-B3 IP (e) and input (f) are shown for each condition. (g-i) PSD-95-ephrin-B3 co-IP from synaptosomes of wild type and Efnb3 -/- animals (g). Control immunoblots indicating the amount of ephrin-B3 IP (h) and input (i) . (j) PSD-95 selectively co-IPs with ephrin-B3 in synaptosomes isolated from P21 wild type brains despite the efficient IP (k) and presence (l) of each of the three ephrin-B f

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 4 Ephrin-B3 regulates subcellular PSD-95 localization (a) Western blots of non-synaptic (S1), crude synaptosomal (P2) and pure synaptosomal (Syn.) fractions from cortices of P10 and P21 wild type and Efnb3 -/- mice probed with the indicated antibodies. Less synaptic enrichment of PSD-95 is observed in synaptosomes from Efnb3 -/- mice. (b) Model and predicted outcomes for the organotypic slice culture experiment. (c-d) Representative two-photon images of cortical pyramidal neurons from wild type and Efnb3 -/- organotypic slices transfected with PSD-95-GFP and the indicated ephrin-B3 rescue constructs. Right panels in c and d are the high contrast representations of tdTomato and PSD-95-GFP. Filled arrowheads indicate PSD-95-GFP localized to a spine; open arrowheads show PSD-95-GFP localized in the dendritic shaft. PSD-95-GFP is diffusely localized in dendritic shafts of Efnb3 -/- neurons (c) and ephrin-B3 shRNA-transfected wild type neurons (d) . Scale bar in c, d : 5 um. (e) The ratio of average GFP pixel intensities in dendritic shafts to the GFP intensities per pixel in puncta was used to measure the fraction of diffuse PSD-95-GFP in c-d (ANOVA, F (4, 93) = 7.155, p-values: wild type// Efnb3 -/- : 0.0105, wild type// Efnb3 -/- + S332D: 0.0233, Efnb3 -/- // Efnb3 -/- + wt-eB3: 0.0285, Efnb3 -/- // Efnb3 -/- + S332A: 0.0015, Efnb3 -/- + wt-eB3// Efnb3 -/- + S332D: 0.0487, Efnb3 -/- + S332A// Efnb3 -/- + S332D: 0.0027; wild type (n = 22), Efnb3 -/- (n = 31), Efnb3 -/- +

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 3 ERK phosphorylation of S332 regulates subcellular localization of ephrin-B3 (a) Schematic representation of the experiments. (b) Representative immunoblots of lysates from DIV14 rat cortical neurons treated with TTX (1uM, 4 hr.) and U0126 (10uM, 4 hr.) to block activity and ERK activation, respectively. Depolarization with KCl (55mM, 1 hr.) was used to elevate ERK activity. ( c ) Ratio of pS332 to ephrin-B3 signals was used to quantify the effect of neuronal activity on S332 phosphorylation (ANOVA, F (3, 12) = 19.27, Tukey's post-hoc, TTX-KCl: p = 0.0012, TTX+U0126-KCl: p

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Fig. 3 Ephrin-B3 is down-regulated in the excited DG region under voluntary running. ( A ) 3D reconstruction of confocal images of rNSCs and GCs in Nestin-GFP mice that were injected with AAV-CaMKII-mCherry. The confocal Z stack images were simulated to 3D images with LAS X software. The arrowheads indicate the direct contact areas of rNSCs and GCs. ( B ) Results of GO Enrichment analysis on the set of 1802 DEGs. The length of each bar indicates the log 10 ( P value), and the vertical axis shows significantly enriched terms. ( C ) The selected nine genes with differential expression are shown in the heat map. ( D ) mRNA from DG excited by the chemogenetic approach was used to detect differential expression of genes selected from previous sequencing data after running ( 28 ). ( E ) The expression of ephrin-B3 in DG tissue from control and hM3Dq mice was detected by Western blot. ( F ) The expression of ephrin-B3 in DG tissue from control and hM4Di mice under voluntary running treatment was detected by Western blot. Results are presented as means +- SEM. * P < 0.05; *** P < 0.001; **** P < 0.0001.

34-3600

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