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Sox5, supplied by Developmental Studies Hybridoma Bank, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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gene exp sox5 mm01264584 m1  (Thermo Fisher)
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Thermo Fisher gene exp sox5 mm01264584 m1
(A, B) Generation of <t>Sox5</t> null (A) and Sox5 icKO (B) mice. (C) Confocal images showing Sox5 expression in dorsal DG of Control and Sox5 null mice by P5 and Sox5 + cells expressing GFAP in Control P5 DG. (D) Confocal images showing GFAP, Sox2, and MCM2 immunostaining in the SGZ (space between white lines) of P5 Control and Sox5 null mice. (E) Quantitation of NSCs/mm 2 (rGFAP + Sox2 + cells) and proliferative MCM2 + NSCs (rGFAP + Sox2 + MCM2 + % over total NSCs) in the SGZ of Control and Sox5 null P5 mice. (F) Confocal images showing GFAP, Sox2, and BrdU immunostaining in P30 Control and Sox5 null mice after P3 BrdU pulse. (G) Scheme of BrdU-long labeling retention experiment. Quantitation of the % of BrdU-L + cells in NSCs (rGFAP + Sox2 + ) in P30 Control and Sox5 null mice. (H) Confocal images showing TdTom, GFAP, and EdU immunostaining in the DG of P30 Control and Sox5 icKO mice. (I) Scheme of EdU-long labeling retention experiment in TAM injected conditional Sox5 icKO mice (P3 > P30). Quantitation of the % of EdU + cells in TdTom + NSCs of P30 Control and Sox5 icKO mice. Data represent mean values ± SEM. * p < 0.05, ** p < 0.01, and *** p < 0.001 by unpaired Student t test. Horizontal dotted lines delineate the SGZ. Scale bars represent 100 µm (C) and 50 µm (D, F, H). Data are available in as a part of Supporting information. Drawings were created through SciDraw.
Gene Exp Sox5 Mm01264584 M1, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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p atf2  (Proteintech)
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Proteintech p atf2
(A, B) Generation of <t>Sox5</t> null (A) and Sox5 icKO (B) mice. (C) Confocal images showing Sox5 expression in dorsal DG of Control and Sox5 null mice by P5 and Sox5 + cells expressing GFAP in Control P5 DG. (D) Confocal images showing GFAP, Sox2, and MCM2 immunostaining in the SGZ (space between white lines) of P5 Control and Sox5 null mice. (E) Quantitation of NSCs/mm 2 (rGFAP + Sox2 + cells) and proliferative MCM2 + NSCs (rGFAP + Sox2 + MCM2 + % over total NSCs) in the SGZ of Control and Sox5 null P5 mice. (F) Confocal images showing GFAP, Sox2, and BrdU immunostaining in P30 Control and Sox5 null mice after P3 BrdU pulse. (G) Scheme of BrdU-long labeling retention experiment. Quantitation of the % of BrdU-L + cells in NSCs (rGFAP + Sox2 + ) in P30 Control and Sox5 null mice. (H) Confocal images showing TdTom, GFAP, and EdU immunostaining in the DG of P30 Control and Sox5 icKO mice. (I) Scheme of EdU-long labeling retention experiment in TAM injected conditional Sox5 icKO mice (P3 > P30). Quantitation of the % of EdU + cells in TdTom + NSCs of P30 Control and Sox5 icKO mice. Data represent mean values ± SEM. * p < 0.05, ** p < 0.01, and *** p < 0.001 by unpaired Student t test. Horizontal dotted lines delineate the SGZ. Scale bars represent 100 µm (C) and 50 µm (D, F, H). Data are available in as a part of Supporting information. Drawings were created through SciDraw.
P Atf2, supplied by Proteintech, 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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sox5  (Proteintech)
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Proteintech sox5
(A, B) Generation of <t>Sox5</t> null (A) and Sox5 icKO (B) mice. (C) Confocal images showing Sox5 expression in dorsal DG of Control and Sox5 null mice by P5 and Sox5 + cells expressing GFAP in Control P5 DG. (D) Confocal images showing GFAP, Sox2, and MCM2 immunostaining in the SGZ (space between white lines) of P5 Control and Sox5 null mice. (E) Quantitation of NSCs/mm 2 (rGFAP + Sox2 + cells) and proliferative MCM2 + NSCs (rGFAP + Sox2 + MCM2 + % over total NSCs) in the SGZ of Control and Sox5 null P5 mice. (F) Confocal images showing GFAP, Sox2, and BrdU immunostaining in P30 Control and Sox5 null mice after P3 BrdU pulse. (G) Scheme of BrdU-long labeling retention experiment. Quantitation of the % of BrdU-L + cells in NSCs (rGFAP + Sox2 + ) in P30 Control and Sox5 null mice. (H) Confocal images showing TdTom, GFAP, and EdU immunostaining in the DG of P30 Control and Sox5 icKO mice. (I) Scheme of EdU-long labeling retention experiment in TAM injected conditional Sox5 icKO mice (P3 > P30). Quantitation of the % of EdU + cells in TdTom + NSCs of P30 Control and Sox5 icKO mice. Data represent mean values ± SEM. * p < 0.05, ** p < 0.01, and *** p < 0.001 by unpaired Student t test. Horizontal dotted lines delineate the SGZ. Scale bars represent 100 µm (C) and 50 µm (D, F, H). Data are available in as a part of Supporting information. Drawings were created through SciDraw.
Sox5, supplied by Proteintech, 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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anti sox5  (Proteintech)
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Proteintech anti sox5
(A, B) Generation of <t>Sox5</t> null (A) and Sox5 icKO (B) mice. (C) Confocal images showing Sox5 expression in dorsal DG of Control and Sox5 null mice by P5 and Sox5 + cells expressing GFAP in Control P5 DG. (D) Confocal images showing GFAP, Sox2, and MCM2 immunostaining in the SGZ (space between white lines) of P5 Control and Sox5 null mice. (E) Quantitation of NSCs/mm 2 (rGFAP + Sox2 + cells) and proliferative MCM2 + NSCs (rGFAP + Sox2 + MCM2 + % over total NSCs) in the SGZ of Control and Sox5 null P5 mice. (F) Confocal images showing GFAP, Sox2, and BrdU immunostaining in P30 Control and Sox5 null mice after P3 BrdU pulse. (G) Scheme of BrdU-long labeling retention experiment. Quantitation of the % of BrdU-L + cells in NSCs (rGFAP + Sox2 + ) in P30 Control and Sox5 null mice. (H) Confocal images showing TdTom, GFAP, and EdU immunostaining in the DG of P30 Control and Sox5 icKO mice. (I) Scheme of EdU-long labeling retention experiment in TAM injected conditional Sox5 icKO mice (P3 > P30). Quantitation of the % of EdU + cells in TdTom + NSCs of P30 Control and Sox5 icKO mice. Data represent mean values ± SEM. * p < 0.05, ** p < 0.01, and *** p < 0.001 by unpaired Student t test. Horizontal dotted lines delineate the SGZ. Scale bars represent 100 µm (C) and 50 µm (D, F, H). Data are available in as a part of Supporting information. Drawings were created through SciDraw.
Anti Sox5, supplied by Proteintech, 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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protein capture reagents program  (Developmental Studies Hybridoma Bank)
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Developmental Studies Hybridoma Bank protein capture reagents program
(A, B) Generation of <t>Sox5</t> null (A) and Sox5 icKO (B) mice. (C) Confocal images showing Sox5 expression in dorsal DG of Control and Sox5 null mice by P5 and Sox5 + cells expressing GFAP in Control P5 DG. (D) Confocal images showing GFAP, Sox2, and MCM2 immunostaining in the SGZ (space between white lines) of P5 Control and Sox5 null mice. (E) Quantitation of NSCs/mm 2 (rGFAP + Sox2 + cells) and proliferative MCM2 + NSCs (rGFAP + Sox2 + MCM2 + % over total NSCs) in the SGZ of Control and Sox5 null P5 mice. (F) Confocal images showing GFAP, Sox2, and BrdU immunostaining in P30 Control and Sox5 null mice after P3 BrdU pulse. (G) Scheme of BrdU-long labeling retention experiment. Quantitation of the % of BrdU-L + cells in NSCs (rGFAP + Sox2 + ) in P30 Control and Sox5 null mice. (H) Confocal images showing TdTom, GFAP, and EdU immunostaining in the DG of P30 Control and Sox5 icKO mice. (I) Scheme of EdU-long labeling retention experiment in TAM injected conditional Sox5 icKO mice (P3 > P30). Quantitation of the % of EdU + cells in TdTom + NSCs of P30 Control and Sox5 icKO mice. Data represent mean values ± SEM. * p < 0.05, ** p < 0.01, and *** p < 0.001 by unpaired Student t test. Horizontal dotted lines delineate the SGZ. Scale bars represent 100 µm (C) and 50 µm (D, F, H). Data are available in as a part of Supporting information. Drawings were created through SciDraw.
Protein Capture Reagents Program, supplied by Developmental Studies Hybridoma Bank, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/protein capture reagents program/product/Developmental Studies Hybridoma Bank
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pcrp sox5 1e3  (Developmental Studies Hybridoma Bank)
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Developmental Studies Hybridoma Bank pcrp sox5 1e3
(A, B) Generation of <t>Sox5</t> null (A) and Sox5 icKO (B) mice. (C) Confocal images showing Sox5 expression in dorsal DG of Control and Sox5 null mice by P5 and Sox5 + cells expressing GFAP in Control P5 DG. (D) Confocal images showing GFAP, Sox2, and MCM2 immunostaining in the SGZ (space between white lines) of P5 Control and Sox5 null mice. (E) Quantitation of NSCs/mm 2 (rGFAP + Sox2 + cells) and proliferative MCM2 + NSCs (rGFAP + Sox2 + MCM2 + % over total NSCs) in the SGZ of Control and Sox5 null P5 mice. (F) Confocal images showing GFAP, Sox2, and BrdU immunostaining in P30 Control and Sox5 null mice after P3 BrdU pulse. (G) Scheme of BrdU-long labeling retention experiment. Quantitation of the % of BrdU-L + cells in NSCs (rGFAP + Sox2 + ) in P30 Control and Sox5 null mice. (H) Confocal images showing TdTom, GFAP, and EdU immunostaining in the DG of P30 Control and Sox5 icKO mice. (I) Scheme of EdU-long labeling retention experiment in TAM injected conditional Sox5 icKO mice (P3 > P30). Quantitation of the % of EdU + cells in TdTom + NSCs of P30 Control and Sox5 icKO mice. Data represent mean values ± SEM. * p < 0.05, ** p < 0.01, and *** p < 0.001 by unpaired Student t test. Horizontal dotted lines delineate the SGZ. Scale bars represent 100 µm (C) and 50 µm (D, F, H). Data are available in as a part of Supporting information. Drawings were created through SciDraw.
Pcrp Sox5 1e3, supplied by Developmental Studies Hybridoma Bank, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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(A, B) Generation of Sox5 null (A) and Sox5 icKO (B) mice. (C) Confocal images showing Sox5 expression in dorsal DG of Control and Sox5 null mice by P5 and Sox5 + cells expressing GFAP in Control P5 DG. (D) Confocal images showing GFAP, Sox2, and MCM2 immunostaining in the SGZ (space between white lines) of P5 Control and Sox5 null mice. (E) Quantitation of NSCs/mm 2 (rGFAP + Sox2 + cells) and proliferative MCM2 + NSCs (rGFAP + Sox2 + MCM2 + % over total NSCs) in the SGZ of Control and Sox5 null P5 mice. (F) Confocal images showing GFAP, Sox2, and BrdU immunostaining in P30 Control and Sox5 null mice after P3 BrdU pulse. (G) Scheme of BrdU-long labeling retention experiment. Quantitation of the % of BrdU-L + cells in NSCs (rGFAP + Sox2 + ) in P30 Control and Sox5 null mice. (H) Confocal images showing TdTom, GFAP, and EdU immunostaining in the DG of P30 Control and Sox5 icKO mice. (I) Scheme of EdU-long labeling retention experiment in TAM injected conditional Sox5 icKO mice (P3 > P30). Quantitation of the % of EdU + cells in TdTom + NSCs of P30 Control and Sox5 icKO mice. Data represent mean values ± SEM. * p < 0.05, ** p < 0.01, and *** p < 0.001 by unpaired Student t test. Horizontal dotted lines delineate the SGZ. Scale bars represent 100 µm (C) and 50 µm (D, F, H). Data are available in as a part of Supporting information. Drawings were created through SciDraw.

Journal: PLOS Biology

Article Title: Sox5 controls the establishment of quiescence in neural stem cells during postnatal development

doi: 10.1371/journal.pbio.3002654

Figure Lengend Snippet: (A, B) Generation of Sox5 null (A) and Sox5 icKO (B) mice. (C) Confocal images showing Sox5 expression in dorsal DG of Control and Sox5 null mice by P5 and Sox5 + cells expressing GFAP in Control P5 DG. (D) Confocal images showing GFAP, Sox2, and MCM2 immunostaining in the SGZ (space between white lines) of P5 Control and Sox5 null mice. (E) Quantitation of NSCs/mm 2 (rGFAP + Sox2 + cells) and proliferative MCM2 + NSCs (rGFAP + Sox2 + MCM2 + % over total NSCs) in the SGZ of Control and Sox5 null P5 mice. (F) Confocal images showing GFAP, Sox2, and BrdU immunostaining in P30 Control and Sox5 null mice after P3 BrdU pulse. (G) Scheme of BrdU-long labeling retention experiment. Quantitation of the % of BrdU-L + cells in NSCs (rGFAP + Sox2 + ) in P30 Control and Sox5 null mice. (H) Confocal images showing TdTom, GFAP, and EdU immunostaining in the DG of P30 Control and Sox5 icKO mice. (I) Scheme of EdU-long labeling retention experiment in TAM injected conditional Sox5 icKO mice (P3 > P30). Quantitation of the % of EdU + cells in TdTom + NSCs of P30 Control and Sox5 icKO mice. Data represent mean values ± SEM. * p < 0.05, ** p < 0.01, and *** p < 0.001 by unpaired Student t test. Horizontal dotted lines delineate the SGZ. Scale bars represent 100 µm (C) and 50 µm (D, F, H). Data are available in as a part of Supporting information. Drawings were created through SciDraw.

Article Snippet: The following Taqman assays probes were used: Anapc (Mm00614339_m1), Axin2 (Mm00443610_m1), Bmp2 (Mm01340178_m1), Bmp3 (Mm00557790_m1), Bmp7 (Mm00432102_m1), Bmpr1b (Mm03023971_m1), Ccna2 (Mm00438063_m1), Ccnb1 (Mm03053893_gH), Ctdsp1 (Mm00778482_s1), Ctdsp2 (Mm01254394_m1), Fstl5 (Mm00618418_m1), Gapdh (Mm99999915_g1), Huwe1 (Mm00615533_m1), Id4 (Mm00499701_m1), Mcm4 (Mm00725863_s1), Myc (Mm00487804_m1), Ppm1a (Mm00725963_s1), Ppm1h (Mm00620945_m1), Smurf2 (Mm03024086_m1), Sox5 (Mm01264584_m1) and Ythdf2 (Mm00661925_m1).

Techniques: Expressing, Control, Immunostaining, Quantitation Assay, Labeling, Injection

(A) Confocal images showing GFAP, Sox2, and MCM2 immunostaining in the SGZ (space between white lines) of P14 Control and Sox5 null mice. (B) Quantitation of NSCs/mm 2 (rGFAP + Sox2 + cells) and proliferative MCM2 + NSCs (% over total NSCs) in Control and Sox5 null P14 mice. (C) Confocal images showing GFAP, Sox2, and BrdU immunostaining in P30 Control and Sox5 null mice after P14 BrdU pulse. (D) Scheme of BrdU-long labeling retention experiment. Quantitation of the % of BrdU-L + cells in NSCs (rGFAP + Sox2 + ) of P30 Control and Sox5 null mice. (E) Confocal images showing TdTom, GFAP, and EdU immunostaining in the SGZ of P30 Control and Sox5 icKO mice. (F) Scheme of EdU-long labeling retention experiment in TAM injected conditional Sox5 icKO mice (P14 > P30). Quantitation of the % of EdU-L + cells in TdTom + NSCs of P30 Control and Sox5 icKO mice. In all graphs, data represent mean value ± SEM. * p < 0.05, ** p < 0.01, and *** p < 0.001 by unpaired Student t test. Scale bars represent 50 µm (A, C) and 20 µm (E). Data are available in as a part of Supporting information. Drawing was created through SciDraw.

Journal: PLOS Biology

Article Title: Sox5 controls the establishment of quiescence in neural stem cells during postnatal development

doi: 10.1371/journal.pbio.3002654

Figure Lengend Snippet: (A) Confocal images showing GFAP, Sox2, and MCM2 immunostaining in the SGZ (space between white lines) of P14 Control and Sox5 null mice. (B) Quantitation of NSCs/mm 2 (rGFAP + Sox2 + cells) and proliferative MCM2 + NSCs (% over total NSCs) in Control and Sox5 null P14 mice. (C) Confocal images showing GFAP, Sox2, and BrdU immunostaining in P30 Control and Sox5 null mice after P14 BrdU pulse. (D) Scheme of BrdU-long labeling retention experiment. Quantitation of the % of BrdU-L + cells in NSCs (rGFAP + Sox2 + ) of P30 Control and Sox5 null mice. (E) Confocal images showing TdTom, GFAP, and EdU immunostaining in the SGZ of P30 Control and Sox5 icKO mice. (F) Scheme of EdU-long labeling retention experiment in TAM injected conditional Sox5 icKO mice (P14 > P30). Quantitation of the % of EdU-L + cells in TdTom + NSCs of P30 Control and Sox5 icKO mice. In all graphs, data represent mean value ± SEM. * p < 0.05, ** p < 0.01, and *** p < 0.001 by unpaired Student t test. Scale bars represent 50 µm (A, C) and 20 µm (E). Data are available in as a part of Supporting information. Drawing was created through SciDraw.

Article Snippet: The following Taqman assays probes were used: Anapc (Mm00614339_m1), Axin2 (Mm00443610_m1), Bmp2 (Mm01340178_m1), Bmp3 (Mm00557790_m1), Bmp7 (Mm00432102_m1), Bmpr1b (Mm03023971_m1), Ccna2 (Mm00438063_m1), Ccnb1 (Mm03053893_gH), Ctdsp1 (Mm00778482_s1), Ctdsp2 (Mm01254394_m1), Fstl5 (Mm00618418_m1), Gapdh (Mm99999915_g1), Huwe1 (Mm00615533_m1), Id4 (Mm00499701_m1), Mcm4 (Mm00725863_s1), Myc (Mm00487804_m1), Ppm1a (Mm00725963_s1), Ppm1h (Mm00620945_m1), Smurf2 (Mm03024086_m1), Sox5 (Mm01264584_m1) and Ythdf2 (Mm00661925_m1).

Techniques: Immunostaining, Control, Quantitation Assay, Labeling, Injection

(A) Confocal images showing GFAP, Sox2, and MCM2 immunostaining in the SGZ of P30 Control and Sox5 null mice. (B) Quantitation of NSCs/mm 2 (rGFAP + Sox2 + cells), and proliferative MCM2 + NSCs (% over total NSCs) of Control and Sox5 null P30 mice. (C) Scheme of BrdU-long labeling retention experiment. Quantitation of the % of BrdU-L + cells in NSCs (rGFAP + Sox2 + ) of P30 Control and Sox5 null mice. (D) Confocal images showing TdTom, GFAP, and MCM2 immunostaining in the SGZ of P30 Control and Sox5 icKO mice. (E) Scheme of Sox5 inducible deletion at P3 > P30 and at P14 > P30 in Sox5 icKO mice by TAM injection. Quantitation of MCM2 + aNSCs in TdTom + rGFAP NSCs of P30 Control and Sox5 icKO mice. (F) Confocal images showing TdTom expression and Nestin, Ki67, and EdU immunostaining in the SGZ of P30 Control mice after the procedure described in (G). (G) Experimental scheme of EdU label retention experiment in P5 > P30 in Sox5 icKO mice to label all shallow/resting and aNSCs (EdU + ) during the P30 > P45 time period. EdU − NSCs could correspond to dormant qNSCs. Quantitation of the indicated subpopulation in the shallow/resting qNSC EdU + TdTom + population (left) or in the dormant EdU - TdTom + NSC population of P45 Control ( n = 5) and Sox5 icKO ( n = 3) mice. In all graphs, data represent mean values ± SEM. * p < 0.05, ** p < 0.01, and *** p < 0.001 by unpaired Student t test. Scale bars represent 50 µm (A and D) and 10 µm in F. Data are available in as a part of Suppor t ing information. Drawings were created through SciDraw.

Journal: PLOS Biology

Article Title: Sox5 controls the establishment of quiescence in neural stem cells during postnatal development

doi: 10.1371/journal.pbio.3002654

Figure Lengend Snippet: (A) Confocal images showing GFAP, Sox2, and MCM2 immunostaining in the SGZ of P30 Control and Sox5 null mice. (B) Quantitation of NSCs/mm 2 (rGFAP + Sox2 + cells), and proliferative MCM2 + NSCs (% over total NSCs) of Control and Sox5 null P30 mice. (C) Scheme of BrdU-long labeling retention experiment. Quantitation of the % of BrdU-L + cells in NSCs (rGFAP + Sox2 + ) of P30 Control and Sox5 null mice. (D) Confocal images showing TdTom, GFAP, and MCM2 immunostaining in the SGZ of P30 Control and Sox5 icKO mice. (E) Scheme of Sox5 inducible deletion at P3 > P30 and at P14 > P30 in Sox5 icKO mice by TAM injection. Quantitation of MCM2 + aNSCs in TdTom + rGFAP NSCs of P30 Control and Sox5 icKO mice. (F) Confocal images showing TdTom expression and Nestin, Ki67, and EdU immunostaining in the SGZ of P30 Control mice after the procedure described in (G). (G) Experimental scheme of EdU label retention experiment in P5 > P30 in Sox5 icKO mice to label all shallow/resting and aNSCs (EdU + ) during the P30 > P45 time period. EdU − NSCs could correspond to dormant qNSCs. Quantitation of the indicated subpopulation in the shallow/resting qNSC EdU + TdTom + population (left) or in the dormant EdU - TdTom + NSC population of P45 Control ( n = 5) and Sox5 icKO ( n = 3) mice. In all graphs, data represent mean values ± SEM. * p < 0.05, ** p < 0.01, and *** p < 0.001 by unpaired Student t test. Scale bars represent 50 µm (A and D) and 10 µm in F. Data are available in as a part of Suppor t ing information. Drawings were created through SciDraw.

Article Snippet: The following Taqman assays probes were used: Anapc (Mm00614339_m1), Axin2 (Mm00443610_m1), Bmp2 (Mm01340178_m1), Bmp3 (Mm00557790_m1), Bmp7 (Mm00432102_m1), Bmpr1b (Mm03023971_m1), Ccna2 (Mm00438063_m1), Ccnb1 (Mm03053893_gH), Ctdsp1 (Mm00778482_s1), Ctdsp2 (Mm01254394_m1), Fstl5 (Mm00618418_m1), Gapdh (Mm99999915_g1), Huwe1 (Mm00615533_m1), Id4 (Mm00499701_m1), Mcm4 (Mm00725863_s1), Myc (Mm00487804_m1), Ppm1a (Mm00725963_s1), Ppm1h (Mm00620945_m1), Smurf2 (Mm03024086_m1), Sox5 (Mm01264584_m1) and Ythdf2 (Mm00661925_m1).

Techniques: Immunostaining, Control, Quantitation Assay, Labeling, Injection, Expressing

(A) Confocal images showing DCX immunostaining in the SGZ of DG at the indicated stages in Control and Sox5 null mice. (B) Time-course quantitation of DCX + cells/mm 3 at the indicated stages in Control and Sox5 null mice. (C) Confocal images showing TdTom + and DCX + immunostaining in the SGZ of P30 Control and Sox5 icKO mice. (D) Scheme of Sox5 inducible deletion at P3 > P30 and at P14 > P30 in Sox5 icKO mice by TAM injection. Quantitation of the % of DCX + cells over total TdTom + cells in P30 Control and Sox5 icKO mice. (E) Confocal images showing GFAP and Sox2 immunostaining in the SGZ of P90 Control and Sox5 null mice. (F) Quantitation of NSCs (rGFAP + Sox2 + cells)/mm 2 at the indicated stages in Control and Sox5 null mice. At least three animals and three sections/animal were analyzed for each immunostaining. In all graphs, data are mean value ± SEM. * p < 0.05, ** p < 0.01, and *** p < 0.001 by unpaired Student t test. Horizontal lines in pictures indica t e the SGZ. Scale bars represent 100 µm (A) and 30 µm (C, E). Data are available in as a part of Supporting information. Drawing was created through SciDraw.

Journal: PLOS Biology

Article Title: Sox5 controls the establishment of quiescence in neural stem cells during postnatal development

doi: 10.1371/journal.pbio.3002654

Figure Lengend Snippet: (A) Confocal images showing DCX immunostaining in the SGZ of DG at the indicated stages in Control and Sox5 null mice. (B) Time-course quantitation of DCX + cells/mm 3 at the indicated stages in Control and Sox5 null mice. (C) Confocal images showing TdTom + and DCX + immunostaining in the SGZ of P30 Control and Sox5 icKO mice. (D) Scheme of Sox5 inducible deletion at P3 > P30 and at P14 > P30 in Sox5 icKO mice by TAM injection. Quantitation of the % of DCX + cells over total TdTom + cells in P30 Control and Sox5 icKO mice. (E) Confocal images showing GFAP and Sox2 immunostaining in the SGZ of P90 Control and Sox5 null mice. (F) Quantitation of NSCs (rGFAP + Sox2 + cells)/mm 2 at the indicated stages in Control and Sox5 null mice. At least three animals and three sections/animal were analyzed for each immunostaining. In all graphs, data are mean value ± SEM. * p < 0.05, ** p < 0.01, and *** p < 0.001 by unpaired Student t test. Horizontal lines in pictures indica t e the SGZ. Scale bars represent 100 µm (A) and 30 µm (C, E). Data are available in as a part of Supporting information. Drawing was created through SciDraw.

Article Snippet: The following Taqman assays probes were used: Anapc (Mm00614339_m1), Axin2 (Mm00443610_m1), Bmp2 (Mm01340178_m1), Bmp3 (Mm00557790_m1), Bmp7 (Mm00432102_m1), Bmpr1b (Mm03023971_m1), Ccna2 (Mm00438063_m1), Ccnb1 (Mm03053893_gH), Ctdsp1 (Mm00778482_s1), Ctdsp2 (Mm01254394_m1), Fstl5 (Mm00618418_m1), Gapdh (Mm99999915_g1), Huwe1 (Mm00615533_m1), Id4 (Mm00499701_m1), Mcm4 (Mm00725863_s1), Myc (Mm00487804_m1), Ppm1a (Mm00725963_s1), Ppm1h (Mm00620945_m1), Smurf2 (Mm03024086_m1), Sox5 (Mm01264584_m1) and Ythdf2 (Mm00661925_m1).

Techniques: Immunostaining, Control, Quantitation Assay, Injection

(A) Experimental approach of RNA-Seq analysis of FACS-sorted NSCs from Control and Sox5 null P14 mice. (B) Principal Component Analysis (PCA) of DEGs indicating independent segregation of the two genotypes analyzed, Control (gray) and Sox5 null (violet). Heat Map of DEGs in Sox5 null vs. Control. Volcano Plot of Log2FC and −log10 (padjusted) of DEGs between Control and Sox5 null mice. Several relevant genes are indicated in boxes, and those related to quiescence ( Foxo3, Foxo4, Huwe1 ) are downregulated, whereas genes related to cell cycle ( Myc ) and BMP canonical signaling ( BMP2, BMP4, BMP5 ) are upregulated in Sox5 null vs. Control DG. (C) Enrichment analysis of GO terms related to Biological Processes and Molecular Functions in DEGs of Sox5 null vs. Control NSCs. (D) KEGG and PANTHER pathways analysis of DEGs of Sox5 null vs. Control NSCs. DEGs were selected for their p -adjusted value < 0.05. (E, F) Node graphs of most representative upregulated cell cycle genes and downregulated TGF-β/BMP Superfamily signaling pathway genes generated by the PANEV software (PAthway NEtwork Visualizer; v. 17.0). (G) Heat Map of DEGs manually curated in the list of BMP signaling pathway genes comparing Sox5 null vs. Control NSCs. (H) Quantitation of the relative levels of mRNA expression by quantitative PCR for the indicated transcripts in Sox5 null vs. Control NSCs prepared in vitro (see ). Results are shown as 2 −ΔΔCT normalized with respect to GAPDH mRNA and relative to P14 NSCs Control values (dashed line y-axis = 1). Control and Sox5 null P14 mice ( N = 3 and 3, respectively) were used to prepared DG NSC cultures and were analyzed for each condition and experimental replicates were performed. (I) Scheme of the BMP canonical signaling pathway. Data represent mean values ± SEM. * p < 0.05; ** p < 0.01; *** p < 0.001according to a Student t test. Data are available in as a part of Supporting information. Drawings were created through SciDraw, Bioicons, Bioart, and Inkscape.

Journal: PLOS Biology

Article Title: Sox5 controls the establishment of quiescence in neural stem cells during postnatal development

doi: 10.1371/journal.pbio.3002654

Figure Lengend Snippet: (A) Experimental approach of RNA-Seq analysis of FACS-sorted NSCs from Control and Sox5 null P14 mice. (B) Principal Component Analysis (PCA) of DEGs indicating independent segregation of the two genotypes analyzed, Control (gray) and Sox5 null (violet). Heat Map of DEGs in Sox5 null vs. Control. Volcano Plot of Log2FC and −log10 (padjusted) of DEGs between Control and Sox5 null mice. Several relevant genes are indicated in boxes, and those related to quiescence ( Foxo3, Foxo4, Huwe1 ) are downregulated, whereas genes related to cell cycle ( Myc ) and BMP canonical signaling ( BMP2, BMP4, BMP5 ) are upregulated in Sox5 null vs. Control DG. (C) Enrichment analysis of GO terms related to Biological Processes and Molecular Functions in DEGs of Sox5 null vs. Control NSCs. (D) KEGG and PANTHER pathways analysis of DEGs of Sox5 null vs. Control NSCs. DEGs were selected for their p -adjusted value < 0.05. (E, F) Node graphs of most representative upregulated cell cycle genes and downregulated TGF-β/BMP Superfamily signaling pathway genes generated by the PANEV software (PAthway NEtwork Visualizer; v. 17.0). (G) Heat Map of DEGs manually curated in the list of BMP signaling pathway genes comparing Sox5 null vs. Control NSCs. (H) Quantitation of the relative levels of mRNA expression by quantitative PCR for the indicated transcripts in Sox5 null vs. Control NSCs prepared in vitro (see ). Results are shown as 2 −ΔΔCT normalized with respect to GAPDH mRNA and relative to P14 NSCs Control values (dashed line y-axis = 1). Control and Sox5 null P14 mice ( N = 3 and 3, respectively) were used to prepared DG NSC cultures and were analyzed for each condition and experimental replicates were performed. (I) Scheme of the BMP canonical signaling pathway. Data represent mean values ± SEM. * p < 0.05; ** p < 0.01; *** p < 0.001according to a Student t test. Data are available in as a part of Supporting information. Drawings were created through SciDraw, Bioicons, Bioart, and Inkscape.

Article Snippet: The following Taqman assays probes were used: Anapc (Mm00614339_m1), Axin2 (Mm00443610_m1), Bmp2 (Mm01340178_m1), Bmp3 (Mm00557790_m1), Bmp7 (Mm00432102_m1), Bmpr1b (Mm03023971_m1), Ccna2 (Mm00438063_m1), Ccnb1 (Mm03053893_gH), Ctdsp1 (Mm00778482_s1), Ctdsp2 (Mm01254394_m1), Fstl5 (Mm00618418_m1), Gapdh (Mm99999915_g1), Huwe1 (Mm00615533_m1), Id4 (Mm00499701_m1), Mcm4 (Mm00725863_s1), Myc (Mm00487804_m1), Ppm1a (Mm00725963_s1), Ppm1h (Mm00620945_m1), Smurf2 (Mm03024086_m1), Sox5 (Mm01264584_m1) and Ythdf2 (Mm00661925_m1).

Techniques: RNA Sequencing, Control, Generated, Software, Quantitation Assay, Expressing, Real-time Polymerase Chain Reaction, In Vitro

(A) Methodological approach for NSC culture. (B) Representative bright field images of P14 neurospheres loaded with DFFDA (green) under proliferating (FGF2) and quiescence (FGF2 + BMP4) culture conditions for 6 days. (C) Quantitation using flow cytometry of DFFDA high and DFFDA low cells at the indicated culture condition. (D) Confocal images showing DFFDA fluorescence and Ki67, Id4, or pSmad1/5/9 expression in P14 NSCs. (E) Quantitation of DFFDA high and DFFDA low cells in Control and Sox5 null mice in proliferation. (F) Quantitation of the percentage of pSmad1/5/9 + , Ki67 + , and Id4 + cells among DFFDA high and DFFDA low cells from Control mice cultured in FGF2. (G) Quantitation of the percentage of pSmad1/5/9 + , Ki67 + , and Id4 + cells relative to total NSCs in Control and Sox5 null mice cultured in FGF2 (passages 2–6). (H) Confocal images showing Ki67 and Id4 immunostaining in the indicated conditions. (I) Quantitation of the percentage of Id4 + cells, Id4 + /Ki67 − (qNSCs), and Id4 − /Ki67 + (aNSCs). (J) Quantitation of total Ki67 + cells relative to total cell number in Control and Sox5 null mice under the indicated culture conditions. (K, L) Quantitation of the percentage of pSmad1/5/9 + (K) or Ki67 + (L) cells relative to total NSCs in Control and Sox5 null mice cultured in FGF2 or FGF2 + BMP4, with or without the addition of the BMP signaling inhibitor Noggin (Nog), as indicated. Three independent experiments were performed of control and Sox5 null P14 DG NSCs (2–3 different animal of each). At least 500 cells were analyzed for each condition. Data represent mean values ± SEM. * p < 0.05; ** p < 0.01; *** p < 0.001 according to a Student t test (E–G) and two-way ANOVA test (I–M). Scale bars represent 100 µm (B) and 30 µm (D, H). Data are available in and as a part of Supporting information. Drawings were created through SciDraw, Bioicons and Inkscape.

Journal: PLOS Biology

Article Title: Sox5 controls the establishment of quiescence in neural stem cells during postnatal development

doi: 10.1371/journal.pbio.3002654

Figure Lengend Snippet: (A) Methodological approach for NSC culture. (B) Representative bright field images of P14 neurospheres loaded with DFFDA (green) under proliferating (FGF2) and quiescence (FGF2 + BMP4) culture conditions for 6 days. (C) Quantitation using flow cytometry of DFFDA high and DFFDA low cells at the indicated culture condition. (D) Confocal images showing DFFDA fluorescence and Ki67, Id4, or pSmad1/5/9 expression in P14 NSCs. (E) Quantitation of DFFDA high and DFFDA low cells in Control and Sox5 null mice in proliferation. (F) Quantitation of the percentage of pSmad1/5/9 + , Ki67 + , and Id4 + cells among DFFDA high and DFFDA low cells from Control mice cultured in FGF2. (G) Quantitation of the percentage of pSmad1/5/9 + , Ki67 + , and Id4 + cells relative to total NSCs in Control and Sox5 null mice cultured in FGF2 (passages 2–6). (H) Confocal images showing Ki67 and Id4 immunostaining in the indicated conditions. (I) Quantitation of the percentage of Id4 + cells, Id4 + /Ki67 − (qNSCs), and Id4 − /Ki67 + (aNSCs). (J) Quantitation of total Ki67 + cells relative to total cell number in Control and Sox5 null mice under the indicated culture conditions. (K, L) Quantitation of the percentage of pSmad1/5/9 + (K) or Ki67 + (L) cells relative to total NSCs in Control and Sox5 null mice cultured in FGF2 or FGF2 + BMP4, with or without the addition of the BMP signaling inhibitor Noggin (Nog), as indicated. Three independent experiments were performed of control and Sox5 null P14 DG NSCs (2–3 different animal of each). At least 500 cells were analyzed for each condition. Data represent mean values ± SEM. * p < 0.05; ** p < 0.01; *** p < 0.001 according to a Student t test (E–G) and two-way ANOVA test (I–M). Scale bars represent 100 µm (B) and 30 µm (D, H). Data are available in and as a part of Supporting information. Drawings were created through SciDraw, Bioicons and Inkscape.

Article Snippet: The following Taqman assays probes were used: Anapc (Mm00614339_m1), Axin2 (Mm00443610_m1), Bmp2 (Mm01340178_m1), Bmp3 (Mm00557790_m1), Bmp7 (Mm00432102_m1), Bmpr1b (Mm03023971_m1), Ccna2 (Mm00438063_m1), Ccnb1 (Mm03053893_gH), Ctdsp1 (Mm00778482_s1), Ctdsp2 (Mm01254394_m1), Fstl5 (Mm00618418_m1), Gapdh (Mm99999915_g1), Huwe1 (Mm00615533_m1), Id4 (Mm00499701_m1), Mcm4 (Mm00725863_s1), Myc (Mm00487804_m1), Ppm1a (Mm00725963_s1), Ppm1h (Mm00620945_m1), Smurf2 (Mm03024086_m1), Sox5 (Mm01264584_m1) and Ythdf2 (Mm00661925_m1).

Techniques: Quantitation Assay, Flow Cytometry, Fluorescence, Expressing, Control, Cell Culture, Immunostaining

(A) Confocal images showing Nestin, pSmad1/5/9 and MCM2 immunostaining in the SGZ of P14 Control and Sox5 null mice. (B) Quantitation of pSmad1/5/9 levels intensity (arbitrary units) in aNSCs (MCM2 + ) and qNSCs (MCM2 − ) in Control and Sox5 null P14 mice. (C) Confocal images showing Nestin, Ki67, and Id4 immunostaining in the SGZ of P14 Control and Sox5 null mice. (D) Quantitation of Id4 + Ki67 + and Id4 + Ki67 − Nestin + NSCs per mm 2 in Control and Sox5 null P14 mice. (E) Confocal images showing GFAP, Nestin, Sox2, and MCM2 immunostaining in the SGZ of P28 Control and Sox5 null mice injected with LDN193189 (LDN). (F) Scheme of BMP signaling inhibitor LDN193189 i.p. administration from P12 > P18 in Control and Sox5 null mice. Quantitation of MCM2 + cells in NSCs (Nestin + GFAP + Sox2 + ) in Control and Sox5 null P28 mice injected with LDN or DMSO. Panel left of the right graph is the same as shown in . (G, H) Summary of NSC quiescence dynamics during early postnatal DG development (G) and their alterations in Sox5 null mice (H). Data are available in as a part of Supporting information. Drawings were created through Inkscape.

Journal: PLOS Biology

Article Title: Sox5 controls the establishment of quiescence in neural stem cells during postnatal development

doi: 10.1371/journal.pbio.3002654

Figure Lengend Snippet: (A) Confocal images showing Nestin, pSmad1/5/9 and MCM2 immunostaining in the SGZ of P14 Control and Sox5 null mice. (B) Quantitation of pSmad1/5/9 levels intensity (arbitrary units) in aNSCs (MCM2 + ) and qNSCs (MCM2 − ) in Control and Sox5 null P14 mice. (C) Confocal images showing Nestin, Ki67, and Id4 immunostaining in the SGZ of P14 Control and Sox5 null mice. (D) Quantitation of Id4 + Ki67 + and Id4 + Ki67 − Nestin + NSCs per mm 2 in Control and Sox5 null P14 mice. (E) Confocal images showing GFAP, Nestin, Sox2, and MCM2 immunostaining in the SGZ of P28 Control and Sox5 null mice injected with LDN193189 (LDN). (F) Scheme of BMP signaling inhibitor LDN193189 i.p. administration from P12 > P18 in Control and Sox5 null mice. Quantitation of MCM2 + cells in NSCs (Nestin + GFAP + Sox2 + ) in Control and Sox5 null P28 mice injected with LDN or DMSO. Panel left of the right graph is the same as shown in . (G, H) Summary of NSC quiescence dynamics during early postnatal DG development (G) and their alterations in Sox5 null mice (H). Data are available in as a part of Supporting information. Drawings were created through Inkscape.

Article Snippet: The following Taqman assays probes were used: Anapc (Mm00614339_m1), Axin2 (Mm00443610_m1), Bmp2 (Mm01340178_m1), Bmp3 (Mm00557790_m1), Bmp7 (Mm00432102_m1), Bmpr1b (Mm03023971_m1), Ccna2 (Mm00438063_m1), Ccnb1 (Mm03053893_gH), Ctdsp1 (Mm00778482_s1), Ctdsp2 (Mm01254394_m1), Fstl5 (Mm00618418_m1), Gapdh (Mm99999915_g1), Huwe1 (Mm00615533_m1), Id4 (Mm00499701_m1), Mcm4 (Mm00725863_s1), Myc (Mm00487804_m1), Ppm1a (Mm00725963_s1), Ppm1h (Mm00620945_m1), Smurf2 (Mm03024086_m1), Sox5 (Mm01264584_m1) and Ythdf2 (Mm00661925_m1).

Techniques: Immunostaining, Control, Quantitation Assay, Injection