MA5-15008

Caffa I, Spagnolo V, Vernieri C, Valdemarin F, Becherini P, Wei M, Brandhorst S, Zucal C, Driehuis E, Ferrando L, Piacente F, Tagliafico A, Cilli M, Mastracci L, Vellone VG, Piazza S, Cremonini AL, Gradaschi R, Mantero C, Passalacqua M, Ballestrero A, Zoppoli G, Cea M, Arrighi A, Odetti P, Monacelli F, Salvadori G, Cortellino S, Clevers H, De Braud F, Sukkar SG, Provenzani A, Longo VD, Nencioni A
Nature 2020 Jul;583(7817):620-624

METTL3-mediated N(6)-methyladenosine mRNA modification enhances long-term memory consolidation.

Zhang Z, Wang M, Xie D, Huang Z, Zhang L, Yang Y, Ma D, Li W, Zhou Q, Yang YG, Wang XJ
Cell research 2018 Nov;28(11):1050-1061

Melatonin attenuates scopolamine-induced memory/synaptic disorder by rescuing EPACs/miR-124/Egr1 pathway.

Wang X, Wang ZH, Wu YY, Tang H, Tan L, Wang X, Gao XY, Xiong YS, Liu D, Wang JZ, Zhu LQ
Molecular neurobiology 2013 Feb;47(1):373-81

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of EGR1 in extracts from serum-starved PC12 cells, untreated or NGF-treated (100 ng/mL for 2 hours), using EGR1 monoclonal antibody (Product # MA5-15008).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis was performed on modified whole cell extracts (1% SDS) of PC-12 (Lane 1), PC-12 Serum Starved (Serum starved for overnight) (Lane 2), PC-12 Serum Starved and then treated with NGF (Serum Starved for overnight, NGF treatment at 100ng/ml for 2hr) (Lane 3), NIH/3T3 (Lane 4), SW480 (Lane 5), A-431 (Lane 6), HeLa (Lane 7) and A-375 (Lane 8). The blot was probed with Anti-EGR1 Monoclonal Antibody (Product # MA5-15008, 1:1000 dilution) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.25 µg/ml, 1:4000 dilution). A 56 kDa band corresponding to EGR1 was detected across the cell lines and tissues tested. Also there was an induction of EGR1 in PC-12 cells upon NGF treatment.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescent analysis of EGR1 in dissociated PC12 cells using an EGR1 monoclonal antibody (Product # MA5-15008) (green) and a MEK1/2 monoclonal antibody (red).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Fig. 5 m 6 A promotes the translation of immediate-early genes upon activity induction. a Immediate-early genes (IEGs) are comparably induced by fear conditioning training in the hippocampus of both the CTRL (blue) and cKO (red) mice. b Increment of m 6 A modification abundance on IEG mRNAs after fear conditioning training in hippocampus samples of the CTRL mice but not the Mettl3 cKO mice. IP, immunoprecipitated fraction. c Translation of IEGs is impaired in the cKO mice after training. d Immunofluorescent image of EGR1 in CA1 region before and after fear conditioning training (1 h and 4 h). e Translation of IEGs is impaired in Mettl3 -KO primary cortical neurons, which can be rescued by re-expression of wildtype Mettl3 but not mutated Mettl3 . Student's t -test, * P < 0.05, ** P < 0.01, *** P < 0.001, N.S., not significant, ( a ) n = 2 replicates, ( b ), ( c ) and ( e ) n = 3 replicates

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: FIGURE 8 Expression of 35 key genes and EGR1 in AKI samples and sham samples. (A) Transcriptional levels of 35 important genes including 21 hub genes and 14 other important upregulated genes in our own AKI samples. (B) Translational levels of Egr1 in AKI samples and sham samples. (C) Western blot of EGR1. (D,E) Immunofluorescence of EGR1. * p < 0.05, ** p < 0.01, *** p < 0.001.