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(A) Schematic timeline of brain organoids generation from mESCs. mESCs and brain organoids developed for 7, 14 and 21 days were profiled by transcriptomics and proteomics. Part of the figure was created with Biorender. (B) Immunostaining of mESCs-derived brain organoids. Cultures of mESCs were stained with NANOG (green), <t>POU5F1/OCT4</t> (red), and nuclei with DAPI (blue). Scale bars: 10 µm. (C) Organoid cryosections were stained with NESTIN and PAX6 (at day 7), ELAVL3 and TBR1 (at day 14), and TUBB3 and GFAP (at day 21). Scale bars: 100 µm. (D) PCA showing the distribution of the transcriptomes of mESCs, brain organoids (at days 7, 14, and 21), and NBB samples. (E) Venn diagram showing the overlapping upregulated and downregulated genes between D21 organoids and NBB, both compared to mESCs. (F) Histograms of enriched GO terms in NBB vs ESCs and D21 organoids vs ESCs performed on upregulated genes (black and grey bars) and on downregulated genes (red and light red bars) determined using DAVID. For each term category (Cellular component, molecular function, and biological process), the five highest enriched terms in each gene list are shown.
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(A) Schematic timeline of brain organoids generation from mESCs. mESCs and brain organoids developed for 7, 14 and 21 days were profiled by transcriptomics and proteomics. Part of the figure was created with Biorender. (B) Immunostaining of mESCs-derived brain organoids. Cultures of mESCs were stained with NANOG (green), <t>POU5F1/OCT4</t> (red), and nuclei with DAPI (blue). Scale bars: 10 µm. (C) Organoid cryosections were stained with NESTIN and PAX6 (at day 7), ELAVL3 and TBR1 (at day 14), and TUBB3 and GFAP (at day 21). Scale bars: 100 µm. (D) PCA showing the distribution of the transcriptomes of mESCs, brain organoids (at days 7, 14, and 21), and NBB samples. (E) Venn diagram showing the overlapping upregulated and downregulated genes between D21 organoids and NBB, both compared to mESCs. (F) Histograms of enriched GO terms in NBB vs ESCs and D21 organoids vs ESCs performed on upregulated genes (black and grey bars) and on downregulated genes (red and light red bars) determined using DAVID. For each term category (Cellular component, molecular function, and biological process), the five highest enriched terms in each gene list are shown.
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(A) Schematic timeline of brain organoids generation from mESCs. mESCs and brain organoids developed for 7, 14 and 21 days were profiled by transcriptomics and proteomics. Part of the figure was created with Biorender. (B) Immunostaining of mESCs-derived brain organoids. Cultures of mESCs were stained with NANOG (green), <t>POU5F1/OCT4</t> (red), and nuclei with DAPI (blue). Scale bars: 10 µm. (C) Organoid cryosections were stained with NESTIN and PAX6 (at day 7), ELAVL3 and TBR1 (at day 14), and TUBB3 and GFAP (at day 21). Scale bars: 100 µm. (D) PCA showing the distribution of the transcriptomes of mESCs, brain organoids (at days 7, 14, and 21), and NBB samples. (E) Venn diagram showing the overlapping upregulated and downregulated genes between D21 organoids and NBB, both compared to mESCs. (F) Histograms of enriched GO terms in NBB vs ESCs and D21 organoids vs ESCs performed on upregulated genes (black and grey bars) and on downregulated genes (red and light red bars) determined using DAVID. For each term category (Cellular component, molecular function, and biological process), the five highest enriched terms in each gene list are shown.
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(A) Schematic timeline of brain organoids generation from mESCs. mESCs and brain organoids developed for 7, 14 and 21 days were profiled by transcriptomics and proteomics. Part of the figure was created with Biorender. (B) Immunostaining of mESCs-derived brain organoids. Cultures of mESCs were stained with NANOG (green), <t>POU5F1/OCT4</t> (red), and nuclei with DAPI (blue). Scale bars: 10 µm. (C) Organoid cryosections were stained with NESTIN and PAX6 (at day 7), ELAVL3 and TBR1 (at day 14), and TUBB3 and GFAP (at day 21). Scale bars: 100 µm. (D) PCA showing the distribution of the transcriptomes of mESCs, brain organoids (at days 7, 14, and 21), and NBB samples. (E) Venn diagram showing the overlapping upregulated and downregulated genes between D21 organoids and NBB, both compared to mESCs. (F) Histograms of enriched GO terms in NBB vs ESCs and D21 organoids vs ESCs performed on upregulated genes (black and grey bars) and on downregulated genes (red and light red bars) determined using DAVID. For each term category (Cellular component, molecular function, and biological process), the five highest enriched terms in each gene list are shown.
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(A) Schematic timeline of brain organoids generation from mESCs. mESCs and brain organoids developed for 7, 14 and 21 days were profiled by transcriptomics and proteomics. Part of the figure was created with Biorender. (B) Immunostaining of mESCs-derived brain organoids. Cultures of mESCs were stained with NANOG (green), <t>POU5F1/OCT4</t> (red), and nuclei with DAPI (blue). Scale bars: 10 µm. (C) Organoid cryosections were stained with NESTIN and PAX6 (at day 7), ELAVL3 and TBR1 (at day 14), and TUBB3 and GFAP (at day 21). Scale bars: 100 µm. (D) PCA showing the distribution of the transcriptomes of mESCs, brain organoids (at days 7, 14, and 21), and NBB samples. (E) Venn diagram showing the overlapping upregulated and downregulated genes between D21 organoids and NBB, both compared to mESCs. (F) Histograms of enriched GO terms in NBB vs ESCs and D21 organoids vs ESCs performed on upregulated genes (black and grey bars) and on downregulated genes (red and light red bars) determined using DAVID. For each term category (Cellular component, molecular function, and biological process), the five highest enriched terms in each gene list are shown.
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(A) Schematic timeline of brain organoids generation from mESCs. mESCs and brain organoids developed for 7, 14 and 21 days were profiled by transcriptomics and proteomics. Part of the figure was created with Biorender. (B) Immunostaining of mESCs-derived brain organoids. Cultures of mESCs were stained with NANOG (green), <t>POU5F1/OCT4</t> (red), and nuclei with DAPI (blue). Scale bars: 10 µm. (C) Organoid cryosections were stained with NESTIN and PAX6 (at day 7), ELAVL3 and TBR1 (at day 14), and TUBB3 and GFAP (at day 21). Scale bars: 100 µm. (D) PCA showing the distribution of the transcriptomes of mESCs, brain organoids (at days 7, 14, and 21), and NBB samples. (E) Venn diagram showing the overlapping upregulated and downregulated genes between D21 organoids and NBB, both compared to mESCs. (F) Histograms of enriched GO terms in NBB vs ESCs and D21 organoids vs ESCs performed on upregulated genes (black and grey bars) and on downregulated genes (red and light red bars) determined using DAVID. For each term category (Cellular component, molecular function, and biological process), the five highest enriched terms in each gene list are shown.
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(A) Schematic timeline of brain organoids generation from mESCs. mESCs and brain organoids developed for 7, 14 and 21 days were profiled by transcriptomics and proteomics. Part of the figure was created with Biorender. (B) Immunostaining of mESCs-derived brain organoids. Cultures of mESCs were stained with NANOG (green), <t>POU5F1/OCT4</t> (red), and nuclei with DAPI (blue). Scale bars: 10 µm. (C) Organoid cryosections were stained with NESTIN and PAX6 (at day 7), ELAVL3 and TBR1 (at day 14), and TUBB3 and GFAP (at day 21). Scale bars: 100 µm. (D) PCA showing the distribution of the transcriptomes of mESCs, brain organoids (at days 7, 14, and 21), and NBB samples. (E) Venn diagram showing the overlapping upregulated and downregulated genes between D21 organoids and NBB, both compared to mESCs. (F) Histograms of enriched GO terms in NBB vs ESCs and D21 organoids vs ESCs performed on upregulated genes (black and grey bars) and on downregulated genes (red and light red bars) determined using DAVID. For each term category (Cellular component, molecular function, and biological process), the five highest enriched terms in each gene list are shown.
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(A) Schematic timeline of brain organoids generation from mESCs. mESCs and brain organoids developed for 7, 14 and 21 days were profiled by transcriptomics and proteomics. Part of the figure was created with Biorender. (B) Immunostaining of mESCs-derived brain organoids. Cultures of mESCs were stained with NANOG (green), <t>POU5F1/OCT4</t> (red), and nuclei with DAPI (blue). Scale bars: 10 µm. (C) Organoid cryosections were stained with NESTIN and PAX6 (at day 7), ELAVL3 and TBR1 (at day 14), and TUBB3 and GFAP (at day 21). Scale bars: 100 µm. (D) PCA showing the distribution of the transcriptomes of mESCs, brain organoids (at days 7, 14, and 21), and NBB samples. (E) Venn diagram showing the overlapping upregulated and downregulated genes between D21 organoids and NBB, both compared to mESCs. (F) Histograms of enriched GO terms in NBB vs ESCs and D21 organoids vs ESCs performed on upregulated genes (black and grey bars) and on downregulated genes (red and light red bars) determined using DAVID. For each term category (Cellular component, molecular function, and biological process), the five highest enriched terms in each gene list are shown.
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(A) Schematic timeline of brain organoids generation from mESCs. mESCs and brain organoids developed for 7, 14 and 21 days were profiled by transcriptomics and proteomics. Part of the figure was created with Biorender. (B) Immunostaining of mESCs-derived brain organoids. Cultures of mESCs were stained with NANOG (green), POU5F1/OCT4 (red), and nuclei with DAPI (blue). Scale bars: 10 µm. (C) Organoid cryosections were stained with NESTIN and PAX6 (at day 7), ELAVL3 and TBR1 (at day 14), and TUBB3 and GFAP (at day 21). Scale bars: 100 µm. (D) PCA showing the distribution of the transcriptomes of mESCs, brain organoids (at days 7, 14, and 21), and NBB samples. (E) Venn diagram showing the overlapping upregulated and downregulated genes between D21 organoids and NBB, both compared to mESCs. (F) Histograms of enriched GO terms in NBB vs ESCs and D21 organoids vs ESCs performed on upregulated genes (black and grey bars) and on downregulated genes (red and light red bars) determined using DAVID. For each term category (Cellular component, molecular function, and biological process), the five highest enriched terms in each gene list are shown.

Journal: bioRxiv

Article Title: The RNA and protein landscapes of mouse brain organoids

doi: 10.64898/2026.03.13.711293

Figure Lengend Snippet: (A) Schematic timeline of brain organoids generation from mESCs. mESCs and brain organoids developed for 7, 14 and 21 days were profiled by transcriptomics and proteomics. Part of the figure was created with Biorender. (B) Immunostaining of mESCs-derived brain organoids. Cultures of mESCs were stained with NANOG (green), POU5F1/OCT4 (red), and nuclei with DAPI (blue). Scale bars: 10 µm. (C) Organoid cryosections were stained with NESTIN and PAX6 (at day 7), ELAVL3 and TBR1 (at day 14), and TUBB3 and GFAP (at day 21). Scale bars: 100 µm. (D) PCA showing the distribution of the transcriptomes of mESCs, brain organoids (at days 7, 14, and 21), and NBB samples. (E) Venn diagram showing the overlapping upregulated and downregulated genes between D21 organoids and NBB, both compared to mESCs. (F) Histograms of enriched GO terms in NBB vs ESCs and D21 organoids vs ESCs performed on upregulated genes (black and grey bars) and on downregulated genes (red and light red bars) determined using DAVID. For each term category (Cellular component, molecular function, and biological process), the five highest enriched terms in each gene list are shown.

Article Snippet: The primary antibodies were (species, provider; catalog number): anti-NESTIN –Rat-401- (mouse, Santa-Cruz, sc-33677); PAX6 (Rabbit, Covance, PRB-278P); NANOG (mouse, BD Pharmingen, 560259); POU5F1 (rabbit, Cell Signalling, 2840), ELAVL3 (mouse, Santa Cruz, sc-515624), TUBB3 (mouse, Covance, MMS-435P); TBR1 (rabbit, Cell Signalling, 49661), REELIN (mouse, Covance; MAB5364), GRM5 (rabbit, Millipore, AB5675), and GFAP (rabbit, Dako, Z0334).

Techniques: Transcriptomics, Immunostaining, Derivative Assay, Staining