Journal: Cell reports

Article Title: Integrative analysis identifies region- and sex-specific gene networks and Mef2c as a mediator of anxiety-like behavior.

doi: 10.1016/j.celrep.2024.114455

Figure Lengend Snippet: Figure 7. In vivo down-regulation of Mef2c in CA3 recapitulated transcriptional remodeling in CA3 of male susceptible mice (A) Heatmap of DEGs after in vivo down-regulation of Mef2c in CA3 (FDR <0.05). (B and C) Enriched biological processes (B) and cellular components (C) in down-regulated DEGs after in vivo down-regulation of Mef2c in CA3. (D) Venn diagram of overlap between M-cyan genes and up-regulated and down-regulated DEGs after in vivo down-regulation of Mef2c in CA3. (E) Enrichment in down-regulated DEGs after in vivo down-regulation of Mef2c in CA3 in each network module. (F) Enriched biological processes in overlapping M-cyan genes and down-regulated DEGs after in vivo down-regulation of Mef2c in CA3.

Article Snippet: The sections were then treated as follows: 1, incubation in 0.3% Triton X-100 for 30 min; 2, incubation in 3% bovine serum albumin (BSA) for 30 min; 3, incubation with diluted antibodies rabbit anti-Mef2c (1:100; Proteintech, Chicago, USA), mouse anti-CaMKIIa (1:100; Santa Cruz Biotechnology, Dallas, USA), mouse anti-GABA(1:300; Sigma-Aldrich, St. Louis,USA), overnight at 4 C; 4, washing; 5, incubation with diluted Alexa dyetagged secondary antibodies (1:200; Invitrogen, Karlsbad, USA) for 2 h at room temperature; 6, washing; 7, incubation with DAPI staining solution (Beyotime, Shanghai, China) for 10 min; 8, washing; 9, coverslipping and observation.

Techniques: In Vivo

Journal: Cell reports

Article Title: Integrative analysis identifies region- and sex-specific gene networks and Mef2c as a mediator of anxiety-like behavior.

doi: 10.1016/j.celrep.2024.114455

Figure Lengend Snippet: Figure 7. In vivo down-regulation of Mef2c in CA3 recapitulated transcriptional remodeling in CA3 of male susceptible mice (A) Heatmap of DEGs after in vivo down-regulation of Mef2c in CA3 (FDR <0.05). (B and C) Enriched biological processes (B) and cellular components (C) in down-regulated DEGs after in vivo down-regulation of Mef2c in CA3. (D) Venn diagram of overlap between M-cyan genes and up-regulated and down-regulated DEGs after in vivo down-regulation of Mef2c in CA3. (E) Enrichment in down-regulated DEGs after in vivo down-regulation of Mef2c in CA3 in each network module. (F) Enriched biological processes in overlapping M-cyan genes and down-regulated DEGs after in vivo down-regulation of Mef2c in CA3.

Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Rabbit anti-Mef2c Proteintech Cat#10056-1-AP; RRID: AB_513447 Rabbit anti-Ppp1r9b Biorbyt Cat#orb158484; RRID: AB_3099414 Rabbit anti-GAPDH Huabio Cat#ET1601-4; RRID: AB_3069615 Rabbit lgG Cell Signaling Technology Cat# 3900; RRID: AB_1550038 Mouse anti-Plxna2 Santa Cruz Cat#Sc-393939; RRID: AB_3099415 Mouse anti-GABA Sigma-Aldrich Cat# A0310; RRID: AB_476667 Mouse anti-CaMKIIa Santa Cruz Cat# sc-13141; RRID: AB_3220744 Alexa Fluor 488-anti-mouse secondary antibody Invitrogen Cat# A21202; RRID: AB_141607 Alexa Fluor 594-anti-rabbit secondary antibody Invitrogen Cat# A11055; RRID: AB_2534102 Bacterial and viral strains AAV2/9-CaMKIIa-mCherry-50miR-30ashRNA(mef2c)-30miR-30a-WPREs BrainVTA, Wuhan Cat# PT-8223 AAV2/9-CaMKIIa-mCherry-50miR-30ashRNA(scramble)-30miR-30a-WPREs BrainVTA, Wuhan Cat# PT-2485 AAV2/9-CaMKIIa-Mef2c-P2A-mCherryWPRE-hGH pA BrainVTA, Wuhan Cat# PT-8664 AAV2/9-CaMKIIa-mCherry -WPRE-hGH pA BrainVTA, Wuhan Cat# PT-0108 AAV2/9-hysn-mCherry-50miR-30ashRNA(Ppp1r9b)-30miR-30a-WPREs BrainVTA, Wuhan Cat# PT-10086 AAV2/9-hysn-mCherry-50miR-30ashRNA(scramble)-30miR-30a-WPREs BrainVTA, Wuhan Cat# PT-2321 AAV2/9-CaMKIIa-hM3D(Gq)-mCherryWPREs BrainVTA, Wuhan Cat# PT-0049 AAV2/9-CaMKIIa-hM4D(Gi)-mCherryWPREs BrainVTA, Wuhan Cat# PT-0017 Chemicals Clozapine N-oxide (CNO) Sigma-Aldrich Cat# C0832 Deposited data RNA Sequencing Data This paper GEO: GSE266128 GSE254448 Experimental models: Organisms/strains Mouse: C57BL/6J Beijing Vital River Laboratory Animal Technology Co., Ltd. (China) N/A Oligonucleotides See STAR Methods for qPCR primer sequences This paper N/A Software and algorithms GraphPad Prism 8 GraphPad software https://www.graphpad.com/ SPSS (v17.0) SPSS software https://www.ibm.com/spss Clampex Molecular Devices https://www.moleculardevices.com/ products/axon-patch-clamp-system Clampfit v11.1 Molecular Devices https://www.moleculardevices.com/ products/axon-patch-clamp-system Any-Maze Stoelting http://www.anymaze.co.uk/index.htm (Continued on next page) 18 Cell Reports 43, 114455, July 23, 2024

Techniques: In Vivo