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gene exp en2 mm00438710 m1  (Thermo Fisher)
85
Thermo Fisher gene exp en2 mm00438710 m1
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)
93
Cusabio en2
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.
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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gene exp en2 hs00171321 m1  (Thermo Fisher)
86
Thermo Fisher gene exp en2 hs00171321 m1
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 Hs00171321 M1, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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anti-human antibody against en2  (Novus Biologicals)
90
Novus Biologicals anti-human antibody against en2
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.
Anti Human Antibody Against En2, supplied by Novus Biologicals, 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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rabbit anti-en2 antibody  (Novus Biologicals)
90
Novus Biologicals rabbit anti-en2 antibody
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.
Rabbit Anti En2 Antibody, supplied by Novus Biologicals, 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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sirnas specific for en2  (Shanghai GenePharma)
90
Shanghai GenePharma sirnas specific for en2
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.
Sirnas Specific For En2, supplied by Shanghai GenePharma, 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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en2 flox  (Jackson Laboratory)
90
Jackson Laboratory en2 flox
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)
86
Danaher Inc anti en2 antibody
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.
Anti En2 Antibody, supplied by Danaher Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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goat anti-en2  (Santa Cruz Biotechnology)
90
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