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en2  (Cusabio)


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    Cusabio en2
    En2, supplied by Cusabio, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 93 stars, based on 1 article reviews
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    A. Experimental timeline. Pregnant heterozygous mice were injected with LPS or saline at E16, E17, and E18. 4 hours after the last injection, the fetal forebrain was dissected for RNA-seq analysis. B, C. A volcano plot illustrating DEGs in the forebrain of E18. The red dots represent significantly upregulated genes, the blue dots represent significantly downregulated genes (|log2 FC| ≥ 1 and FDR < 0.05), and the black dots represent insignificant differentially expressed genes. (B) DEGs in the forebrain of Lcn2 KO animals at E18 (C) DEGs in the forebrain of WT MIA at E18, 4 hours after the last LPS injection. D, E. Gene ontology enrichment analysis using Metascape based on the genes deregulated (both up- and downregulated). A bar graph showing the top 5 clusters with the highest p-value. E. Heatmap of enriched terms across input differentially expressed gene lists. F. A Venn diagram of unique and overlapping gene expression between Lcn2 KO and MIA groups. G. The heat map of overlapping genes between Lcn2 KO and MIA. H, I. RT-qPCR confirmation of the results obtained with RNAseq analysis for Pax2 (H) and <t>En2</t> (I) genes. mRNA expression levels in the forebrain were measured 4 hours after the final injection of LPS or saline in pregnant dams. (H) Two-way ANOVA: procedure: F (1, 33) = 4.616; p = 0.0391, genotype: F (1, 33) = 8.232; p = 0.0071, and interaction: F (1, 33) = 1.933; p = 0.1737, followed by Fisher’s LSD test. WT ctrl n = 7, WT MIA n = 11; Lcn2 KO n = 8, Lcn2 KO MIA n = 11. (I) Two-way ANOVA: procedure: F (1, 34) = 11.60; p = 0.0017, genotype: F (1, 34) = 4.019; p = 0.0530, and interaction: F (1, 34) = 7.669, p = 0.0090, followed by Fisher’s LSD post hoc test. WT ctrl n = 8, WT MIA n = 10; Lcn2 KO n = 9, Lcn2 KO MIA n = 11. Data are presented as mean ± SEM.
    Gene Exp En2 Mm00438710 M1, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 85/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    en2  (Cusabio)
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    A. Experimental timeline. Pregnant heterozygous mice were injected with LPS or saline at E16, E17, and E18. 4 hours after the last injection, the fetal forebrain was dissected for RNA-seq analysis. B, C. A volcano plot illustrating DEGs in the forebrain of E18. The red dots represent significantly upregulated genes, the blue dots represent significantly downregulated genes (|log2 FC| ≥ 1 and FDR < 0.05), and the black dots represent insignificant differentially expressed genes. (B) DEGs in the forebrain of Lcn2 KO animals at E18 (C) DEGs in the forebrain of WT MIA at E18, 4 hours after the last LPS injection. D, E. Gene ontology enrichment analysis using Metascape based on the genes deregulated (both up- and downregulated). A bar graph showing the top 5 clusters with the highest p-value. E. Heatmap of enriched terms across input differentially expressed gene lists. F. A Venn diagram of unique and overlapping gene expression between Lcn2 KO and MIA groups. G. The heat map of overlapping genes between Lcn2 KO and MIA. H, I. RT-qPCR confirmation of the results obtained with RNAseq analysis for Pax2 (H) and <t>En2</t> (I) genes. mRNA expression levels in the forebrain were measured 4 hours after the final injection of LPS or saline in pregnant dams. (H) Two-way ANOVA: procedure: F (1, 33) = 4.616; p = 0.0391, genotype: F (1, 33) = 8.232; p = 0.0071, and interaction: F (1, 33) = 1.933; p = 0.1737, followed by Fisher’s LSD test. WT ctrl n = 7, WT MIA n = 11; Lcn2 KO n = 8, Lcn2 KO MIA n = 11. (I) Two-way ANOVA: procedure: F (1, 34) = 11.60; p = 0.0017, genotype: F (1, 34) = 4.019; p = 0.0530, and interaction: F (1, 34) = 7.669, p = 0.0090, followed by Fisher’s LSD post hoc test. WT ctrl n = 8, WT MIA n = 10; Lcn2 KO n = 9, Lcn2 KO MIA n = 11. Data are presented as mean ± SEM.
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    A. Experimental timeline. Pregnant heterozygous mice were injected with LPS or saline at E16, E17, and E18. 4 hours after the last injection, the fetal forebrain was dissected for RNA-seq analysis. B, C. A volcano plot illustrating DEGs in the forebrain of E18. The red dots represent significantly upregulated genes, the blue dots represent significantly downregulated genes (|log2 FC| ≥ 1 and FDR < 0.05), and the black dots represent insignificant differentially expressed genes. (B) DEGs in the forebrain of Lcn2 KO animals at E18 (C) DEGs in the forebrain of WT MIA at E18, 4 hours after the last LPS injection. D, E. Gene ontology enrichment analysis using Metascape based on the genes deregulated (both up- and downregulated). A bar graph showing the top 5 clusters with the highest p-value. E. Heatmap of enriched terms across input differentially expressed gene lists. F. A Venn diagram of unique and overlapping gene expression between Lcn2 KO and MIA groups. G. The heat map of overlapping genes between Lcn2 KO and MIA. H, I. RT-qPCR confirmation of the results obtained with RNAseq analysis for Pax2 (H) and <t>En2</t> (I) genes. mRNA expression levels in the forebrain were measured 4 hours after the final injection of LPS or saline in pregnant dams. (H) Two-way ANOVA: procedure: F (1, 33) = 4.616; p = 0.0391, genotype: F (1, 33) = 8.232; p = 0.0071, and interaction: F (1, 33) = 1.933; p = 0.1737, followed by Fisher’s LSD test. WT ctrl n = 7, WT MIA n = 11; Lcn2 KO n = 8, Lcn2 KO MIA n = 11. (I) Two-way ANOVA: procedure: F (1, 34) = 11.60; p = 0.0017, genotype: F (1, 34) = 4.019; p = 0.0530, and interaction: F (1, 34) = 7.669, p = 0.0090, followed by Fisher’s LSD post hoc test. WT ctrl n = 8, WT MIA n = 10; Lcn2 KO n = 9, Lcn2 KO MIA n = 11. Data are presented as mean ± SEM.
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    A. Experimental timeline. Pregnant heterozygous mice were injected with LPS or saline at E16, E17, and E18. 4 hours after the last injection, the fetal forebrain was dissected for RNA-seq analysis. B, C. A volcano plot illustrating DEGs in the forebrain of E18. The red dots represent significantly upregulated genes, the blue dots represent significantly downregulated genes (|log2 FC| ≥ 1 and FDR < 0.05), and the black dots represent insignificant differentially expressed genes. (B) DEGs in the forebrain of Lcn2 KO animals at E18 (C) DEGs in the forebrain of WT MIA at E18, 4 hours after the last LPS injection. D, E. Gene ontology enrichment analysis using Metascape based on the genes deregulated (both up- and downregulated). A bar graph showing the top 5 clusters with the highest p-value. E. Heatmap of enriched terms across input differentially expressed gene lists. F. A Venn diagram of unique and overlapping gene expression between Lcn2 KO and MIA groups. G. The heat map of overlapping genes between Lcn2 KO and MIA. H, I. RT-qPCR confirmation of the results obtained with RNAseq analysis for Pax2 (H) and <t>En2</t> (I) genes. mRNA expression levels in the forebrain were measured 4 hours after the final injection of LPS or saline in pregnant dams. (H) Two-way ANOVA: procedure: F (1, 33) = 4.616; p = 0.0391, genotype: F (1, 33) = 8.232; p = 0.0071, and interaction: F (1, 33) = 1.933; p = 0.1737, followed by Fisher’s LSD test. WT ctrl n = 7, WT MIA n = 11; Lcn2 KO n = 8, Lcn2 KO MIA n = 11. (I) Two-way ANOVA: procedure: F (1, 34) = 11.60; p = 0.0017, genotype: F (1, 34) = 4.019; p = 0.0530, and interaction: F (1, 34) = 7.669, p = 0.0090, followed by Fisher’s LSD post hoc test. WT ctrl n = 8, WT MIA n = 10; Lcn2 KO n = 9, Lcn2 KO MIA n = 11. Data are presented as mean ± SEM.
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    A. Experimental timeline. Pregnant heterozygous mice were injected with LPS or saline at E16, E17, and E18. 4 hours after the last injection, the fetal forebrain was dissected for RNA-seq analysis. B, C. A volcano plot illustrating DEGs in the forebrain of E18. The red dots represent significantly upregulated genes, the blue dots represent significantly downregulated genes (|log2 FC| ≥ 1 and FDR < 0.05), and the black dots represent insignificant differentially expressed genes. (B) DEGs in the forebrain of Lcn2 KO animals at E18 (C) DEGs in the forebrain of WT MIA at E18, 4 hours after the last LPS injection. D, E. Gene ontology enrichment analysis using Metascape based on the genes deregulated (both up- and downregulated). A bar graph showing the top 5 clusters with the highest p-value. E. Heatmap of enriched terms across input differentially expressed gene lists. F. A Venn diagram of unique and overlapping gene expression between Lcn2 KO and MIA groups. G. The heat map of overlapping genes between Lcn2 KO and MIA. H, I. RT-qPCR confirmation of the results obtained with RNAseq analysis for Pax2 (H) and <t>En2</t> (I) genes. mRNA expression levels in the forebrain were measured 4 hours after the final injection of LPS or saline in pregnant dams. (H) Two-way ANOVA: procedure: F (1, 33) = 4.616; p = 0.0391, genotype: F (1, 33) = 8.232; p = 0.0071, and interaction: F (1, 33) = 1.933; p = 0.1737, followed by Fisher’s LSD test. WT ctrl n = 7, WT MIA n = 11; Lcn2 KO n = 8, Lcn2 KO MIA n = 11. (I) Two-way ANOVA: procedure: F (1, 34) = 11.60; p = 0.0017, genotype: F (1, 34) = 4.019; p = 0.0530, and interaction: F (1, 34) = 7.669, p = 0.0090, followed by Fisher’s LSD post hoc test. WT ctrl n = 8, WT MIA n = 10; Lcn2 KO n = 9, Lcn2 KO MIA n = 11. Data are presented as mean ± SEM.
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    sirnas specific for en2  (Shanghai GenePharma)
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    A. Experimental timeline. Pregnant heterozygous mice were injected with LPS or saline at E16, E17, and E18. 4 hours after the last injection, the fetal forebrain was dissected for RNA-seq analysis. B, C. A volcano plot illustrating DEGs in the forebrain of E18. The red dots represent significantly upregulated genes, the blue dots represent significantly downregulated genes (|log2 FC| ≥ 1 and FDR < 0.05), and the black dots represent insignificant differentially expressed genes. (B) DEGs in the forebrain of Lcn2 KO animals at E18 (C) DEGs in the forebrain of WT MIA at E18, 4 hours after the last LPS injection. D, E. Gene ontology enrichment analysis using Metascape based on the genes deregulated (both up- and downregulated). A bar graph showing the top 5 clusters with the highest p-value. E. Heatmap of enriched terms across input differentially expressed gene lists. F. A Venn diagram of unique and overlapping gene expression between Lcn2 KO and MIA groups. G. The heat map of overlapping genes between Lcn2 KO and MIA. H, I. RT-qPCR confirmation of the results obtained with RNAseq analysis for Pax2 (H) and <t>En2</t> (I) genes. mRNA expression levels in the forebrain were measured 4 hours after the final injection of LPS or saline in pregnant dams. (H) Two-way ANOVA: procedure: F (1, 33) = 4.616; p = 0.0391, genotype: F (1, 33) = 8.232; p = 0.0071, and interaction: F (1, 33) = 1.933; p = 0.1737, followed by Fisher’s LSD test. WT ctrl n = 7, WT MIA n = 11; Lcn2 KO n = 8, Lcn2 KO MIA n = 11. (I) Two-way ANOVA: procedure: F (1, 34) = 11.60; p = 0.0017, genotype: F (1, 34) = 4.019; p = 0.0530, and interaction: F (1, 34) = 7.669, p = 0.0090, followed by Fisher’s LSD post hoc test. WT ctrl n = 8, WT MIA n = 10; Lcn2 KO n = 9, Lcn2 KO MIA n = 11. Data are presented as mean ± SEM.
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    a,b, Schematic (left) and representative images (right) of sagittal sections stained for tdTomato (magenta) in SepW1-Cre; Ai75D mice showing recombination at E14.5 ( b ) but not at E13.5 ( a ) in eCN in the NTZ marked by MEIS2 (green). NTZ = nuclear transitory zone. c , Schematic (left) and representative sagittal image (right) of tdTomato (magenta) expression in SepW1-Cre; Ai75D mice showing recombination at postnatal day (P1) in TBR2+ (green) unipolar brush cells. d , Schematic (left) and representative sagittal images (right) of tdTomato (magenta) expression in SepW1-Cre; Ai75D mice showing recombination at P7 in p27+ (green) differentiated granule cells in the internal granule cell layer (IGL), but not in proliferating granule cell precursors in the external granule cell layer (EGL). Scale bars = 100 um; inset scale bars = 20 um. e , Representative images of coronal sections stained for NeuN (single channel), NeuN (green) and tdTomato (magenta) co-labeling in the posterior CN of SepW1-Cre; Ai75D and SepW1-En1/2 CKO; Ai75D mice ( SepW1-Cre/+; En1 <t>flox/flox</t> ; <t>En2</t> flox/flox ; R26 LSL-nls-tdTomato/+ ). NeuN labeling near the MedP of mutants are ectopic unipolar brush cells that are not TBR2+ or MEIS2+ (confirmed in Krishnamurthy et al., 2024). Abbreviations: MedP=Posterior medial; IntP=Posterior interposed. Scale bars for low magnification = 500 um; scale bars for high magnification = 100 um. Scale bars in a , b , c = 250 um; inset scale bars = 50 um.
    En2 Flox, supplied by Jackson Laboratory, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    anti en2 antibody  (Danaher Inc)
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    a,b, Schematic (left) and representative images (right) of sagittal sections stained for tdTomato (magenta) in SepW1-Cre; Ai75D mice showing recombination at E14.5 ( b ) but not at E13.5 ( a ) in eCN in the NTZ marked by MEIS2 (green). NTZ = nuclear transitory zone. c , Schematic (left) and representative sagittal image (right) of tdTomato (magenta) expression in SepW1-Cre; Ai75D mice showing recombination at postnatal day (P1) in TBR2+ (green) unipolar brush cells. d , Schematic (left) and representative sagittal images (right) of tdTomato (magenta) expression in SepW1-Cre; Ai75D mice showing recombination at P7 in p27+ (green) differentiated granule cells in the internal granule cell layer (IGL), but not in proliferating granule cell precursors in the external granule cell layer (EGL). Scale bars = 100 um; inset scale bars = 20 um. e , Representative images of coronal sections stained for NeuN (single channel), NeuN (green) and tdTomato (magenta) co-labeling in the posterior CN of SepW1-Cre; Ai75D and SepW1-En1/2 CKO; Ai75D mice ( SepW1-Cre/+; En1 <t>flox/flox</t> ; <t>En2</t> flox/flox ; R26 LSL-nls-tdTomato/+ ). NeuN labeling near the MedP of mutants are ectopic unipolar brush cells that are not TBR2+ or MEIS2+ (confirmed in Krishnamurthy et al., 2024). Abbreviations: MedP=Posterior medial; IntP=Posterior interposed. Scale bars for low magnification = 500 um; scale bars for high magnification = 100 um. Scale bars in a , b , c = 250 um; inset scale bars = 50 um.
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    Santa Cruz Biotechnology goat anti-en2
    a,b, Schematic (left) and representative images (right) of sagittal sections stained for tdTomato (magenta) in SepW1-Cre; Ai75D mice showing recombination at E14.5 ( b ) but not at E13.5 ( a ) in eCN in the NTZ marked by MEIS2 (green). NTZ = nuclear transitory zone. c , Schematic (left) and representative sagittal image (right) of tdTomato (magenta) expression in SepW1-Cre; Ai75D mice showing recombination at postnatal day (P1) in TBR2+ (green) unipolar brush cells. d , Schematic (left) and representative sagittal images (right) of tdTomato (magenta) expression in SepW1-Cre; Ai75D mice showing recombination at P7 in p27+ (green) differentiated granule cells in the internal granule cell layer (IGL), but not in proliferating granule cell precursors in the external granule cell layer (EGL). Scale bars = 100 um; inset scale bars = 20 um. e , Representative images of coronal sections stained for NeuN (single channel), NeuN (green) and tdTomato (magenta) co-labeling in the posterior CN of SepW1-Cre; Ai75D and SepW1-En1/2 CKO; Ai75D mice ( SepW1-Cre/+; En1 <t>flox/flox</t> ; <t>En2</t> flox/flox ; R26 LSL-nls-tdTomato/+ ). NeuN labeling near the MedP of mutants are ectopic unipolar brush cells that are not TBR2+ or MEIS2+ (confirmed in Krishnamurthy et al., 2024). Abbreviations: MedP=Posterior medial; IntP=Posterior interposed. Scale bars for low magnification = 500 um; scale bars for high magnification = 100 um. Scale bars in a , b , c = 250 um; inset scale bars = 50 um.
    Goat Anti En2, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    A. Experimental timeline. Pregnant heterozygous mice were injected with LPS or saline at E16, E17, and E18. 4 hours after the last injection, the fetal forebrain was dissected for RNA-seq analysis. B, C. A volcano plot illustrating DEGs in the forebrain of E18. The red dots represent significantly upregulated genes, the blue dots represent significantly downregulated genes (|log2 FC| ≥ 1 and FDR < 0.05), and the black dots represent insignificant differentially expressed genes. (B) DEGs in the forebrain of Lcn2 KO animals at E18 (C) DEGs in the forebrain of WT MIA at E18, 4 hours after the last LPS injection. D, E. Gene ontology enrichment analysis using Metascape based on the genes deregulated (both up- and downregulated). A bar graph showing the top 5 clusters with the highest p-value. E. Heatmap of enriched terms across input differentially expressed gene lists. F. A Venn diagram of unique and overlapping gene expression between Lcn2 KO and MIA groups. G. The heat map of overlapping genes between Lcn2 KO and MIA. H, I. RT-qPCR confirmation of the results obtained with RNAseq analysis for Pax2 (H) and En2 (I) genes. mRNA expression levels in the forebrain were measured 4 hours after the final injection of LPS or saline in pregnant dams. (H) Two-way ANOVA: procedure: F (1, 33) = 4.616; p = 0.0391, genotype: F (1, 33) = 8.232; p = 0.0071, and interaction: F (1, 33) = 1.933; p = 0.1737, followed by Fisher’s LSD test. WT ctrl n = 7, WT MIA n = 11; Lcn2 KO n = 8, Lcn2 KO MIA n = 11. (I) Two-way ANOVA: procedure: F (1, 34) = 11.60; p = 0.0017, genotype: F (1, 34) = 4.019; p = 0.0530, and interaction: F (1, 34) = 7.669, p = 0.0090, followed by Fisher’s LSD post hoc test. WT ctrl n = 8, WT MIA n = 10; Lcn2 KO n = 9, Lcn2 KO MIA n = 11. Data are presented as mean ± SEM.

    Journal: bioRxiv

    Article Title: Lcn2 deficiency leads to social impairments independent of maternal immune activation

    doi: 10.1101/2025.06.27.661499

    Figure Lengend Snippet: A. Experimental timeline. Pregnant heterozygous mice were injected with LPS or saline at E16, E17, and E18. 4 hours after the last injection, the fetal forebrain was dissected for RNA-seq analysis. B, C. A volcano plot illustrating DEGs in the forebrain of E18. The red dots represent significantly upregulated genes, the blue dots represent significantly downregulated genes (|log2 FC| ≥ 1 and FDR < 0.05), and the black dots represent insignificant differentially expressed genes. (B) DEGs in the forebrain of Lcn2 KO animals at E18 (C) DEGs in the forebrain of WT MIA at E18, 4 hours after the last LPS injection. D, E. Gene ontology enrichment analysis using Metascape based on the genes deregulated (both up- and downregulated). A bar graph showing the top 5 clusters with the highest p-value. E. Heatmap of enriched terms across input differentially expressed gene lists. F. A Venn diagram of unique and overlapping gene expression between Lcn2 KO and MIA groups. G. The heat map of overlapping genes between Lcn2 KO and MIA. H, I. RT-qPCR confirmation of the results obtained with RNAseq analysis for Pax2 (H) and En2 (I) genes. mRNA expression levels in the forebrain were measured 4 hours after the final injection of LPS or saline in pregnant dams. (H) Two-way ANOVA: procedure: F (1, 33) = 4.616; p = 0.0391, genotype: F (1, 33) = 8.232; p = 0.0071, and interaction: F (1, 33) = 1.933; p = 0.1737, followed by Fisher’s LSD test. WT ctrl n = 7, WT MIA n = 11; Lcn2 KO n = 8, Lcn2 KO MIA n = 11. (I) Two-way ANOVA: procedure: F (1, 34) = 11.60; p = 0.0017, genotype: F (1, 34) = 4.019; p = 0.0530, and interaction: F (1, 34) = 7.669, p = 0.0090, followed by Fisher’s LSD post hoc test. WT ctrl n = 8, WT MIA n = 10; Lcn2 KO n = 9, Lcn2 KO MIA n = 11. Data are presented as mean ± SEM.

    Article Snippet: RNA was reverse transcribed with SuperScript™ IV VILO (# 11766050, Thermo Fisher Scientific, Waltham, MA, USA) according to the manufacturer’s instructions. cDNA was amplified with TaqMan probes (Lcn2 Mm01324470_m1, Gapdh Mm99999915_g1, En2 Mm00438710_m1, Pax2: Mm01217939_m1, ThermoFisher, Waltham, MA, USA) that were specific for the mouse.

    Techniques: Injection, Saline, RNA Sequencing, Gene Expression, Quantitative RT-PCR, Expressing

    a,b, Schematic (left) and representative images (right) of sagittal sections stained for tdTomato (magenta) in SepW1-Cre; Ai75D mice showing recombination at E14.5 ( b ) but not at E13.5 ( a ) in eCN in the NTZ marked by MEIS2 (green). NTZ = nuclear transitory zone. c , Schematic (left) and representative sagittal image (right) of tdTomato (magenta) expression in SepW1-Cre; Ai75D mice showing recombination at postnatal day (P1) in TBR2+ (green) unipolar brush cells. d , Schematic (left) and representative sagittal images (right) of tdTomato (magenta) expression in SepW1-Cre; Ai75D mice showing recombination at P7 in p27+ (green) differentiated granule cells in the internal granule cell layer (IGL), but not in proliferating granule cell precursors in the external granule cell layer (EGL). Scale bars = 100 um; inset scale bars = 20 um. e , Representative images of coronal sections stained for NeuN (single channel), NeuN (green) and tdTomato (magenta) co-labeling in the posterior CN of SepW1-Cre; Ai75D and SepW1-En1/2 CKO; Ai75D mice ( SepW1-Cre/+; En1 flox/flox ; En2 flox/flox ; R26 LSL-nls-tdTomato/+ ). NeuN labeling near the MedP of mutants are ectopic unipolar brush cells that are not TBR2+ or MEIS2+ (confirmed in Krishnamurthy et al., 2024). Abbreviations: MedP=Posterior medial; IntP=Posterior interposed. Scale bars for low magnification = 500 um; scale bars for high magnification = 100 um. Scale bars in a , b , c = 250 um; inset scale bars = 50 um.

    Journal: bioRxiv

    Article Title: Cerebellar output neurons impair non-motor behaviors by altering development of extracerebellar connectivity

    doi: 10.1101/2024.07.08.602496

    Figure Lengend Snippet: a,b, Schematic (left) and representative images (right) of sagittal sections stained for tdTomato (magenta) in SepW1-Cre; Ai75D mice showing recombination at E14.5 ( b ) but not at E13.5 ( a ) in eCN in the NTZ marked by MEIS2 (green). NTZ = nuclear transitory zone. c , Schematic (left) and representative sagittal image (right) of tdTomato (magenta) expression in SepW1-Cre; Ai75D mice showing recombination at postnatal day (P1) in TBR2+ (green) unipolar brush cells. d , Schematic (left) and representative sagittal images (right) of tdTomato (magenta) expression in SepW1-Cre; Ai75D mice showing recombination at P7 in p27+ (green) differentiated granule cells in the internal granule cell layer (IGL), but not in proliferating granule cell precursors in the external granule cell layer (EGL). Scale bars = 100 um; inset scale bars = 20 um. e , Representative images of coronal sections stained for NeuN (single channel), NeuN (green) and tdTomato (magenta) co-labeling in the posterior CN of SepW1-Cre; Ai75D and SepW1-En1/2 CKO; Ai75D mice ( SepW1-Cre/+; En1 flox/flox ; En2 flox/flox ; R26 LSL-nls-tdTomato/+ ). NeuN labeling near the MedP of mutants are ectopic unipolar brush cells that are not TBR2+ or MEIS2+ (confirmed in Krishnamurthy et al., 2024). Abbreviations: MedP=Posterior medial; IntP=Posterior interposed. Scale bars for low magnification = 500 um; scale bars for high magnification = 100 um. Scale bars in a , b , c = 250 um; inset scale bars = 50 um.

    Article Snippet: The following mouse lines were used in the study: Atoh1-Cre (JAX #011104) , tetO-Cre (JAX #006234) , R26 LSL-nls-tdTomato (Ai75D, JAX #025106) , 129S1/SvImJ (JAX #002448), Swiss Webster mice (Taconic Biosciences catalog #SW), En1 flox (JAX #007918) , En2 flox (JAX #008872) , Atoh1-tTA , En1 Cre (JAX #007916) , lgs7 TRE-lox-tdTomato- STOP-lox-DTA*G 128 D (or lgs7 DRAGON-DTA , JAX #034778) , and SepW1-Cre (MMRRC #036190-UCD) .

    Techniques: Staining, Expressing, Labeling

    a , b , Representative images of immunofluorescent staining NeuN (singe channel) and NeuN (green), tdTomato (magenta) co-labeling in of coronal sections of the anterior ( a ) and posterior ( b ) CN of Atoh1-Cre; Ai75D and Atoh1-En1/2 CKO; Ai75D mice. Abbreviations: MedA=Anterior medial; MedP=Posterior medial; IntA=Anterior interposed; IntP=Posterior interposed; Lat=Lateral. Scale bars = 500 um, scale bars for i - iv = 100 um. c , Experimental design for quantifying excitatory and inhibitory CN neurons in Atoh1-Cre; Ai75D ( Atoh1-Cre/+; R26 LSL-nls-tdTomato/+ ) and Atoh1-En1/2 CKO; Ai75D ( Atoh1-Cre/+; En1 flox/flox ; En2 flox/flox ; R26 LSL-nls-tdTomato/+ ) mice. d , Quantification and distribution of excitatory CN neurons in half of the cerebellum (every second section). Two-way ANOVA: main effect of % mediolateral distance (F 19,120 = 31.38, P < 0.0001), genotype (F 1,120 = 400.1, P < 0.0001), and interaction (F 19,120 = 8.830, P < 0.0001); with post hoc two-tailed t-tests with uncorrected Fisher’s LSD for effect of genotype for bin 10-80% (list of t value for each bin: t 120 = 4.029, 7.169, 8.828, 7.267, 6.726, 6.087, 7.103, 4.759, 5.516, 5.122, 7.85, 5.06, 6, 4.169; all P values: P < 0.0001), for bins 80-85% (t 120 = 3.435, P = 0.0008), and no other comparisons (P ≥ 0.05). Abbreviations: Med=medial; Int=interposed; Lat=lateral. e , Quantification and distribution of inhibitory CN neurons in half of the cerebellum (every second section). Two-way ANOVA: main effect of % mediolateral distance (F 19,120 = 23.97, P < 0.0001), and genotype (F 1,120 = 50.81, P < 0.0001), but not interaction (F 19,120 = 1.659, P = 0.0531); with post hoc two-tailed t-tests with uncorrected Fisher’s LSD for effect of genotype for bin 20-25% (t 120 = 2.094, P = 0.0384), bin 55-60% (t 120 = 2.141, P = 0.0343), bin 60-65% (t 120 = 4.114, P < 0.0001), bin 80-85% (t 120 = 5.316, P <0.0001), and no other comparisons (P ≥ 0.05). Abbreviations: Med=medial; Int=interposed; Lat=lateral. Data are presented as mean values ± SEM.

    Journal: bioRxiv

    Article Title: Cerebellar output neurons impair non-motor behaviors by altering development of extracerebellar connectivity

    doi: 10.1101/2024.07.08.602496

    Figure Lengend Snippet: a , b , Representative images of immunofluorescent staining NeuN (singe channel) and NeuN (green), tdTomato (magenta) co-labeling in of coronal sections of the anterior ( a ) and posterior ( b ) CN of Atoh1-Cre; Ai75D and Atoh1-En1/2 CKO; Ai75D mice. Abbreviations: MedA=Anterior medial; MedP=Posterior medial; IntA=Anterior interposed; IntP=Posterior interposed; Lat=Lateral. Scale bars = 500 um, scale bars for i - iv = 100 um. c , Experimental design for quantifying excitatory and inhibitory CN neurons in Atoh1-Cre; Ai75D ( Atoh1-Cre/+; R26 LSL-nls-tdTomato/+ ) and Atoh1-En1/2 CKO; Ai75D ( Atoh1-Cre/+; En1 flox/flox ; En2 flox/flox ; R26 LSL-nls-tdTomato/+ ) mice. d , Quantification and distribution of excitatory CN neurons in half of the cerebellum (every second section). Two-way ANOVA: main effect of % mediolateral distance (F 19,120 = 31.38, P < 0.0001), genotype (F 1,120 = 400.1, P < 0.0001), and interaction (F 19,120 = 8.830, P < 0.0001); with post hoc two-tailed t-tests with uncorrected Fisher’s LSD for effect of genotype for bin 10-80% (list of t value for each bin: t 120 = 4.029, 7.169, 8.828, 7.267, 6.726, 6.087, 7.103, 4.759, 5.516, 5.122, 7.85, 5.06, 6, 4.169; all P values: P < 0.0001), for bins 80-85% (t 120 = 3.435, P = 0.0008), and no other comparisons (P ≥ 0.05). Abbreviations: Med=medial; Int=interposed; Lat=lateral. e , Quantification and distribution of inhibitory CN neurons in half of the cerebellum (every second section). Two-way ANOVA: main effect of % mediolateral distance (F 19,120 = 23.97, P < 0.0001), and genotype (F 1,120 = 50.81, P < 0.0001), but not interaction (F 19,120 = 1.659, P = 0.0531); with post hoc two-tailed t-tests with uncorrected Fisher’s LSD for effect of genotype for bin 20-25% (t 120 = 2.094, P = 0.0384), bin 55-60% (t 120 = 2.141, P = 0.0343), bin 60-65% (t 120 = 4.114, P < 0.0001), bin 80-85% (t 120 = 5.316, P <0.0001), and no other comparisons (P ≥ 0.05). Abbreviations: Med=medial; Int=interposed; Lat=lateral. Data are presented as mean values ± SEM.

    Article Snippet: The following mouse lines were used in the study: Atoh1-Cre (JAX #011104) , tetO-Cre (JAX #006234) , R26 LSL-nls-tdTomato (Ai75D, JAX #025106) , 129S1/SvImJ (JAX #002448), Swiss Webster mice (Taconic Biosciences catalog #SW), En1 flox (JAX #007918) , En2 flox (JAX #008872) , Atoh1-tTA , En1 Cre (JAX #007916) , lgs7 TRE-lox-tdTomato- STOP-lox-DTA*G 128 D (or lgs7 DRAGON-DTA , JAX #034778) , and SepW1-Cre (MMRRC #036190-UCD) .

    Techniques: Staining, Labeling, Two Tailed Test