Image data of Evf2RNP Protein Pools_Condensates_Sox2_Smc1a_Smc3_Nono

Description

In Cajigas et al. 2021, we show that Sox2 forms large subnuclear domains as part of the Evf2 long non coding RNA ribonucleoprotein complex (RNP) in E13.5 ganglionic eminences (mouse embryonic forebrain single nuclei). We call these protein pools (PPs). We analyze nuclei from wildtype mice (Evf2+/+), Evf2 lacking (Evf2TS/TS) and Evf1 lacking (Evf1TS/TS, expressing Evf2-5'). Sox2 PPs co-localize with Evf2 RNA clouds at specific DNA sites: Dlx5/6 ultraconserved enhancer, and Evf2 repressed target genes (Akr1b8 and Rbm28). Images are probed with antisense RNA to detect Evf2 RNA clouds, DNA probes to detect Dlx5/6, Akr1b8 or Rbm28, and anti Sox2 antibody to detect Sox2 PPs. Z-stacks are obtained using Zeiss 880 and Zen software. 3D reconstructions are performed using IMARIS software. In the follow-up manuscript, we include analysis of 4 additional Evf2RNPs (Nono, Smarcc2, Smc1a and Smc3) that also form protein pools (protein pool condensates (PPCs)) that colocalize with Sox2 PPs. Evf2-RNP PPCs like Sox2 PPs colocalize with Evf2 DNA targets (Akr1b8, Rbm28) and Dlx5/6; Evf2RNP PPC sizes are regulated by Evf2, as indicated by comparing Evf2 and Evf1 mutants with wildtypes (wildtype mice (Evf2+/+), Evf2 lacking (Evf2TS/TS) and Evf1 lacking (Evf1TS/TS, expressing Evf2-5').

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