recombinant proteins shh r d systems Search Results


recombinant shh  (R&D Systems)
94
R&D Systems recombinant shh
a, Ang1 ISH in transverse sections from E10.5 to E13.5 mouse embryos at brachial level (top). Combined Ang1 ISH (pseudo-colored in red) with Olig2 immunofluorescence (bottom). Arrowheads point to Ang1+ co-localization with Olig2+ pMN-NpCs. Scale bar, 50 μm (bottom) and 100 μm (top). b, plot showing that Ang1 expression (% of the area of Ang1+ signal in ventral NpC domains; left y axis) sequentially overlaps with OpC specification (OpC counts; right y axis) at the pMN between E9.5 and E13.5 (see Methods for more details). Graph shows mean ± s.e.m. (Olig2 counts: E9.5: n = 5 images from one embryo, E10.5: n = 20 images from three independent embryos, E11.5: n = 16 images from two independent embryos, E12.5: n = 24 images from four independent embryos, E13.5: n = 15 images from four independent embryos; Ang1 expression: E9.5: n = 6, E10.5: n = 8, E11.5: n = 7, E12.5: n = 14, E13.5: n = 7 images from two independent embryos). c, d, Relative Ptch (c) and Ang1 (d) expression in NSps derived from E11.5 SC progenitors stimulated with <t>recombinant</t> Shh (500 ng ml−1) and/or Shh inhibitor Sant1 (150 nM). Graph shows mean ± s.e.m. (n = 5 independent experiments; one-way ANOVA. Ptch: Control versus Shh *P = 0.0391; Shh versus Shh+Sant1 *P = 0.0142. Ang1: Control versus Shh *P = 0.0375; Shh versus Shh+Sant1 *P = 0.0242). e, Whole-mount SC explants immunostained for Olig2 and Sox10 to detect OpCs (arrowheads) in the VZ and MZ after culturing in control (vehicle) or recombinant Tie2–Fc (2 μg ml−1) in the y–x plane of Z-stack images. On the right side of each image, an orthogonal projection is shown for the z–y plane of a Z-stack. Scale bar, 100 μm. Open arrowheads indicate OpCs in the pMN (VZ) and filled arrowheads in the MZ. f, Scheme summarizing the phenotype of explants treated with recombinant Tie2–Fc or in control conditions. The z–y–x planes are also depicted to illustrate the positioning of the cells. A scheme of explant preparation is also provided in Extended Data Fig. 2e. g, Quantification of the number of OpCs in the MZ (Olig2+) of Tie2–Fc-treated explants, normalized to vehicle-treated explants. Graph shows mean ± s.e.m. (control n = 13, Tie2–Fc n = 17; 5 independent litters). Unpaired two-sided t-test, *P < 0.0248.
Recombinant Shh, supplied by R&D Systems, 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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human recombinant shh  (R&D Systems)
90
R&D Systems human recombinant shh
a, Ang1 ISH in transverse sections from E10.5 to E13.5 mouse embryos at brachial level (top). Combined Ang1 ISH (pseudo-colored in red) with Olig2 immunofluorescence (bottom). Arrowheads point to Ang1+ co-localization with Olig2+ pMN-NpCs. Scale bar, 50 μm (bottom) and 100 μm (top). b, plot showing that Ang1 expression (% of the area of Ang1+ signal in ventral NpC domains; left y axis) sequentially overlaps with OpC specification (OpC counts; right y axis) at the pMN between E9.5 and E13.5 (see Methods for more details). Graph shows mean ± s.e.m. (Olig2 counts: E9.5: n = 5 images from one embryo, E10.5: n = 20 images from three independent embryos, E11.5: n = 16 images from two independent embryos, E12.5: n = 24 images from four independent embryos, E13.5: n = 15 images from four independent embryos; Ang1 expression: E9.5: n = 6, E10.5: n = 8, E11.5: n = 7, E12.5: n = 14, E13.5: n = 7 images from two independent embryos). c, d, Relative Ptch (c) and Ang1 (d) expression in NSps derived from E11.5 SC progenitors stimulated with <t>recombinant</t> Shh (500 ng ml−1) and/or Shh inhibitor Sant1 (150 nM). Graph shows mean ± s.e.m. (n = 5 independent experiments; one-way ANOVA. Ptch: Control versus Shh *P = 0.0391; Shh versus Shh+Sant1 *P = 0.0142. Ang1: Control versus Shh *P = 0.0375; Shh versus Shh+Sant1 *P = 0.0242). e, Whole-mount SC explants immunostained for Olig2 and Sox10 to detect OpCs (arrowheads) in the VZ and MZ after culturing in control (vehicle) or recombinant Tie2–Fc (2 μg ml−1) in the y–x plane of Z-stack images. On the right side of each image, an orthogonal projection is shown for the z–y plane of a Z-stack. Scale bar, 100 μm. Open arrowheads indicate OpCs in the pMN (VZ) and filled arrowheads in the MZ. f, Scheme summarizing the phenotype of explants treated with recombinant Tie2–Fc or in control conditions. The z–y–x planes are also depicted to illustrate the positioning of the cells. A scheme of explant preparation is also provided in Extended Data Fig. 2e. g, Quantification of the number of OpCs in the MZ (Olig2+) of Tie2–Fc-treated explants, normalized to vehicle-treated explants. Graph shows mean ± s.e.m. (control n = 13, Tie2–Fc n = 17; 5 independent litters). Unpaired two-sided t-test, *P < 0.0248.
Human Recombinant Shh, supplied by R&D Systems, 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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recombinant human shh  (R&D Systems)
93
R&D Systems recombinant human shh
a, Ang1 ISH in transverse sections from E10.5 to E13.5 mouse embryos at brachial level (top). Combined Ang1 ISH (pseudo-colored in red) with Olig2 immunofluorescence (bottom). Arrowheads point to Ang1+ co-localization with Olig2+ pMN-NpCs. Scale bar, 50 μm (bottom) and 100 μm (top). b, plot showing that Ang1 expression (% of the area of Ang1+ signal in ventral NpC domains; left y axis) sequentially overlaps with OpC specification (OpC counts; right y axis) at the pMN between E9.5 and E13.5 (see Methods for more details). Graph shows mean ± s.e.m. (Olig2 counts: E9.5: n = 5 images from one embryo, E10.5: n = 20 images from three independent embryos, E11.5: n = 16 images from two independent embryos, E12.5: n = 24 images from four independent embryos, E13.5: n = 15 images from four independent embryos; Ang1 expression: E9.5: n = 6, E10.5: n = 8, E11.5: n = 7, E12.5: n = 14, E13.5: n = 7 images from two independent embryos). c, d, Relative Ptch (c) and Ang1 (d) expression in NSps derived from E11.5 SC progenitors stimulated with <t>recombinant</t> Shh (500 ng ml−1) and/or Shh inhibitor Sant1 (150 nM). Graph shows mean ± s.e.m. (n = 5 independent experiments; one-way ANOVA. Ptch: Control versus Shh *P = 0.0391; Shh versus Shh+Sant1 *P = 0.0142. Ang1: Control versus Shh *P = 0.0375; Shh versus Shh+Sant1 *P = 0.0242). e, Whole-mount SC explants immunostained for Olig2 and Sox10 to detect OpCs (arrowheads) in the VZ and MZ after culturing in control (vehicle) or recombinant Tie2–Fc (2 μg ml−1) in the y–x plane of Z-stack images. On the right side of each image, an orthogonal projection is shown for the z–y plane of a Z-stack. Scale bar, 100 μm. Open arrowheads indicate OpCs in the pMN (VZ) and filled arrowheads in the MZ. f, Scheme summarizing the phenotype of explants treated with recombinant Tie2–Fc or in control conditions. The z–y–x planes are also depicted to illustrate the positioning of the cells. A scheme of explant preparation is also provided in Extended Data Fig. 2e. g, Quantification of the number of OpCs in the MZ (Olig2+) of Tie2–Fc-treated explants, normalized to vehicle-treated explants. Graph shows mean ± s.e.m. (control n = 13, Tie2–Fc n = 17; 5 independent litters). Unpaired two-sided t-test, *P < 0.0248.
Recombinant Human Shh, supplied by R&D Systems, 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
recombinant human shh - by Bioz Stars, 2026-03
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human shh  (R&D Systems)
94
R&D Systems human shh
a, Ang1 ISH in transverse sections from E10.5 to E13.5 mouse embryos at brachial level (top). Combined Ang1 ISH (pseudo-colored in red) with Olig2 immunofluorescence (bottom). Arrowheads point to Ang1+ co-localization with Olig2+ pMN-NpCs. Scale bar, 50 μm (bottom) and 100 μm (top). b, plot showing that Ang1 expression (% of the area of Ang1+ signal in ventral NpC domains; left y axis) sequentially overlaps with OpC specification (OpC counts; right y axis) at the pMN between E9.5 and E13.5 (see Methods for more details). Graph shows mean ± s.e.m. (Olig2 counts: E9.5: n = 5 images from one embryo, E10.5: n = 20 images from three independent embryos, E11.5: n = 16 images from two independent embryos, E12.5: n = 24 images from four independent embryos, E13.5: n = 15 images from four independent embryos; Ang1 expression: E9.5: n = 6, E10.5: n = 8, E11.5: n = 7, E12.5: n = 14, E13.5: n = 7 images from two independent embryos). c, d, Relative Ptch (c) and Ang1 (d) expression in NSps derived from E11.5 SC progenitors stimulated with <t>recombinant</t> Shh (500 ng ml−1) and/or Shh inhibitor Sant1 (150 nM). Graph shows mean ± s.e.m. (n = 5 independent experiments; one-way ANOVA. Ptch: Control versus Shh *P = 0.0391; Shh versus Shh+Sant1 *P = 0.0142. Ang1: Control versus Shh *P = 0.0375; Shh versus Shh+Sant1 *P = 0.0242). e, Whole-mount SC explants immunostained for Olig2 and Sox10 to detect OpCs (arrowheads) in the VZ and MZ after culturing in control (vehicle) or recombinant Tie2–Fc (2 μg ml−1) in the y–x plane of Z-stack images. On the right side of each image, an orthogonal projection is shown for the z–y plane of a Z-stack. Scale bar, 100 μm. Open arrowheads indicate OpCs in the pMN (VZ) and filled arrowheads in the MZ. f, Scheme summarizing the phenotype of explants treated with recombinant Tie2–Fc or in control conditions. The z–y–x planes are also depicted to illustrate the positioning of the cells. A scheme of explant preparation is also provided in Extended Data Fig. 2e. g, Quantification of the number of OpCs in the MZ (Olig2+) of Tie2–Fc-treated explants, normalized to vehicle-treated explants. Graph shows mean ± s.e.m. (control n = 13, Tie2–Fc n = 17; 5 independent litters). Unpaired two-sided t-test, *P < 0.0248.
Human Shh, supplied by R&D Systems, 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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recombinant human sonic hedgehog  (R&D Systems)
94
R&D Systems recombinant human sonic hedgehog
Parkin and SLP-2 interact in human control fibroblasts and hiPSC-derived neurons. A Representative PLA staining for fibroblasts of a control individual under normal culture conditions and after CCCP treatment (3 h, 10 µM). The PLA signal is visualized in red, DAPI-stained nuclei are shown in blue. Scale bar: 10 µm. Quantification of PLA dots per cell shows a significantly higher PLA signal after CCCP exposure, indicating an increased interaction between the two proteins. Statistical differences were calculated by unpaired Mann Whitney U test ****p ≤ 0.0001. B hiPSC-derived neurons of a control individual were processed using PLA under normal culture conditions and after CCCP treatment (3 h, 10 µM). Scale bar: 10 µm. Quantification of PLA dots per cell shows a significantly higher PLA signal after CCCP exposure. Statistical differences were calculated by unpaired Mann Whitney U test. **p ≤ 0.005. The specificity of the PLA interaction results was confirmed by performing the experiments with only one of the two primary antibodies. C Far-Western blot analysis using <t>recombinant</t> SLP-2 (aa 41–356) and Parkin proteins (aa 1–465 and aa 138–465) shows the direct binding of the two proteins. D Quantification of PLA dots per cell in control and PRKN mutant fibroblast lines under normal culture conditions and after CCCP treatment (3 h, 10 µM) using an antibody directed against the Parkin N-terminus and an anti-SLP-2 antibody. Only in the control lines, CCCP treatment resulted in a significantly increased interaction. Statistical differences were calculated by one-way ANOVA followed by Tukey’s post hoc test to correct for multiple comparisons. * ≤ 0.05, **p ≤ 0.01, ****p ≤ 0.0001
Recombinant Human Sonic Hedgehog, supplied by R&D Systems, 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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recombinant mouse protein  (R&D Systems)
95
R&D Systems recombinant mouse protein
Parkin and SLP-2 interact in human control fibroblasts and hiPSC-derived neurons. A Representative PLA staining for fibroblasts of a control individual under normal culture conditions and after CCCP treatment (3 h, 10 µM). The PLA signal is visualized in red, DAPI-stained nuclei are shown in blue. Scale bar: 10 µm. Quantification of PLA dots per cell shows a significantly higher PLA signal after CCCP exposure, indicating an increased interaction between the two proteins. Statistical differences were calculated by unpaired Mann Whitney U test ****p ≤ 0.0001. B hiPSC-derived neurons of a control individual were processed using PLA under normal culture conditions and after CCCP treatment (3 h, 10 µM). Scale bar: 10 µm. Quantification of PLA dots per cell shows a significantly higher PLA signal after CCCP exposure. Statistical differences were calculated by unpaired Mann Whitney U test. **p ≤ 0.005. The specificity of the PLA interaction results was confirmed by performing the experiments with only one of the two primary antibodies. C Far-Western blot analysis using <t>recombinant</t> SLP-2 (aa 41–356) and Parkin proteins (aa 1–465 and aa 138–465) shows the direct binding of the two proteins. D Quantification of PLA dots per cell in control and PRKN mutant fibroblast lines under normal culture conditions and after CCCP treatment (3 h, 10 µM) using an antibody directed against the Parkin N-terminus and an anti-SLP-2 antibody. Only in the control lines, CCCP treatment resulted in a significantly increased interaction. Statistical differences were calculated by one-way ANOVA followed by Tukey’s post hoc test to correct for multiple comparisons. * ≤ 0.05, **p ≤ 0.01, ****p ≤ 0.0001
Recombinant Mouse Protein, supplied by R&D Systems, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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human sonic93 hedgehog  (R&D Systems)
95
R&D Systems human sonic93 hedgehog
Parkin and SLP-2 interact in human control fibroblasts and hiPSC-derived neurons. A Representative PLA staining for fibroblasts of a control individual under normal culture conditions and after CCCP treatment (3 h, 10 µM). The PLA signal is visualized in red, DAPI-stained nuclei are shown in blue. Scale bar: 10 µm. Quantification of PLA dots per cell shows a significantly higher PLA signal after CCCP exposure, indicating an increased interaction between the two proteins. Statistical differences were calculated by unpaired Mann Whitney U test ****p ≤ 0.0001. B hiPSC-derived neurons of a control individual were processed using PLA under normal culture conditions and after CCCP treatment (3 h, 10 µM). Scale bar: 10 µm. Quantification of PLA dots per cell shows a significantly higher PLA signal after CCCP exposure. Statistical differences were calculated by unpaired Mann Whitney U test. **p ≤ 0.005. The specificity of the PLA interaction results was confirmed by performing the experiments with only one of the two primary antibodies. C Far-Western blot analysis using <t>recombinant</t> SLP-2 (aa 41–356) and Parkin proteins (aa 1–465 and aa 138–465) shows the direct binding of the two proteins. D Quantification of PLA dots per cell in control and PRKN mutant fibroblast lines under normal culture conditions and after CCCP treatment (3 h, 10 µM) using an antibody directed against the Parkin N-terminus and an anti-SLP-2 antibody. Only in the control lines, CCCP treatment resulted in a significantly increased interaction. Statistical differences were calculated by one-way ANOVA followed by Tukey’s post hoc test to correct for multiple comparisons. * ≤ 0.05, **p ≤ 0.01, ****p ≤ 0.0001
Human Sonic93 Hedgehog, supplied by R&D Systems, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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shh  (R&D Systems)
94
R&D Systems shh
Parkin and SLP-2 interact in human control fibroblasts and hiPSC-derived neurons. A Representative PLA staining for fibroblasts of a control individual under normal culture conditions and after CCCP treatment (3 h, 10 µM). The PLA signal is visualized in red, DAPI-stained nuclei are shown in blue. Scale bar: 10 µm. Quantification of PLA dots per cell shows a significantly higher PLA signal after CCCP exposure, indicating an increased interaction between the two proteins. Statistical differences were calculated by unpaired Mann Whitney U test ****p ≤ 0.0001. B hiPSC-derived neurons of a control individual were processed using PLA under normal culture conditions and after CCCP treatment (3 h, 10 µM). Scale bar: 10 µm. Quantification of PLA dots per cell shows a significantly higher PLA signal after CCCP exposure. Statistical differences were calculated by unpaired Mann Whitney U test. **p ≤ 0.005. The specificity of the PLA interaction results was confirmed by performing the experiments with only one of the two primary antibodies. C Far-Western blot analysis using <t>recombinant</t> SLP-2 (aa 41–356) and Parkin proteins (aa 1–465 and aa 138–465) shows the direct binding of the two proteins. D Quantification of PLA dots per cell in control and PRKN mutant fibroblast lines under normal culture conditions and after CCCP treatment (3 h, 10 µM) using an antibody directed against the Parkin N-terminus and an anti-SLP-2 antibody. Only in the control lines, CCCP treatment resulted in a significantly increased interaction. Statistical differences were calculated by one-way ANOVA followed by Tukey’s post hoc test to correct for multiple comparisons. * ≤ 0.05, **p ≤ 0.01, ****p ≤ 0.0001
Shh, supplied by R&D Systems, 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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Average 94 stars, based on 1 article reviews
shh - by Bioz Stars, 2026-03
94/100 stars
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sonic hedgehog (shh  (R&D Systems)
90
R&D Systems sonic hedgehog (shh
Parkin and SLP-2 interact in human control fibroblasts and hiPSC-derived neurons. A Representative PLA staining for fibroblasts of a control individual under normal culture conditions and after CCCP treatment (3 h, 10 µM). The PLA signal is visualized in red, DAPI-stained nuclei are shown in blue. Scale bar: 10 µm. Quantification of PLA dots per cell shows a significantly higher PLA signal after CCCP exposure, indicating an increased interaction between the two proteins. Statistical differences were calculated by unpaired Mann Whitney U test ****p ≤ 0.0001. B hiPSC-derived neurons of a control individual were processed using PLA under normal culture conditions and after CCCP treatment (3 h, 10 µM). Scale bar: 10 µm. Quantification of PLA dots per cell shows a significantly higher PLA signal after CCCP exposure. Statistical differences were calculated by unpaired Mann Whitney U test. **p ≤ 0.005. The specificity of the PLA interaction results was confirmed by performing the experiments with only one of the two primary antibodies. C Far-Western blot analysis using <t>recombinant</t> SLP-2 (aa 41–356) and Parkin proteins (aa 1–465 and aa 138–465) shows the direct binding of the two proteins. D Quantification of PLA dots per cell in control and PRKN mutant fibroblast lines under normal culture conditions and after CCCP treatment (3 h, 10 µM) using an antibody directed against the Parkin N-terminus and an anti-SLP-2 antibody. Only in the control lines, CCCP treatment resulted in a significantly increased interaction. Statistical differences were calculated by one-way ANOVA followed by Tukey’s post hoc test to correct for multiple comparisons. * ≤ 0.05, **p ≤ 0.01, ****p ≤ 0.0001
Sonic Hedgehog (Shh, supplied by R&D Systems, 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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sonic hedgehog  (R&D Systems)
93
R&D Systems sonic hedgehog
Parkin and SLP-2 interact in human control fibroblasts and hiPSC-derived neurons. A Representative PLA staining for fibroblasts of a control individual under normal culture conditions and after CCCP treatment (3 h, 10 µM). The PLA signal is visualized in red, DAPI-stained nuclei are shown in blue. Scale bar: 10 µm. Quantification of PLA dots per cell shows a significantly higher PLA signal after CCCP exposure, indicating an increased interaction between the two proteins. Statistical differences were calculated by unpaired Mann Whitney U test ****p ≤ 0.0001. B hiPSC-derived neurons of a control individual were processed using PLA under normal culture conditions and after CCCP treatment (3 h, 10 µM). Scale bar: 10 µm. Quantification of PLA dots per cell shows a significantly higher PLA signal after CCCP exposure. Statistical differences were calculated by unpaired Mann Whitney U test. **p ≤ 0.005. The specificity of the PLA interaction results was confirmed by performing the experiments with only one of the two primary antibodies. C Far-Western blot analysis using <t>recombinant</t> SLP-2 (aa 41–356) and Parkin proteins (aa 1–465 and aa 138–465) shows the direct binding of the two proteins. D Quantification of PLA dots per cell in control and PRKN mutant fibroblast lines under normal culture conditions and after CCCP treatment (3 h, 10 µM) using an antibody directed against the Parkin N-terminus and an anti-SLP-2 antibody. Only in the control lines, CCCP treatment resulted in a significantly increased interaction. Statistical differences were calculated by one-way ANOVA followed by Tukey’s post hoc test to correct for multiple comparisons. * ≤ 0.05, **p ≤ 0.01, ****p ≤ 0.0001
Sonic Hedgehog, supplied by R&D Systems, 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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Image Search Results


a, Ang1 ISH in transverse sections from E10.5 to E13.5 mouse embryos at brachial level (top). Combined Ang1 ISH (pseudo-colored in red) with Olig2 immunofluorescence (bottom). Arrowheads point to Ang1+ co-localization with Olig2+ pMN-NpCs. Scale bar, 50 μm (bottom) and 100 μm (top). b, plot showing that Ang1 expression (% of the area of Ang1+ signal in ventral NpC domains; left y axis) sequentially overlaps with OpC specification (OpC counts; right y axis) at the pMN between E9.5 and E13.5 (see Methods for more details). Graph shows mean ± s.e.m. (Olig2 counts: E9.5: n = 5 images from one embryo, E10.5: n = 20 images from three independent embryos, E11.5: n = 16 images from two independent embryos, E12.5: n = 24 images from four independent embryos, E13.5: n = 15 images from four independent embryos; Ang1 expression: E9.5: n = 6, E10.5: n = 8, E11.5: n = 7, E12.5: n = 14, E13.5: n = 7 images from two independent embryos). c, d, Relative Ptch (c) and Ang1 (d) expression in NSps derived from E11.5 SC progenitors stimulated with recombinant Shh (500 ng ml−1) and/or Shh inhibitor Sant1 (150 nM). Graph shows mean ± s.e.m. (n = 5 independent experiments; one-way ANOVA. Ptch: Control versus Shh *P = 0.0391; Shh versus Shh+Sant1 *P = 0.0142. Ang1: Control versus Shh *P = 0.0375; Shh versus Shh+Sant1 *P = 0.0242). e, Whole-mount SC explants immunostained for Olig2 and Sox10 to detect OpCs (arrowheads) in the VZ and MZ after culturing in control (vehicle) or recombinant Tie2–Fc (2 μg ml−1) in the y–x plane of Z-stack images. On the right side of each image, an orthogonal projection is shown for the z–y plane of a Z-stack. Scale bar, 100 μm. Open arrowheads indicate OpCs in the pMN (VZ) and filled arrowheads in the MZ. f, Scheme summarizing the phenotype of explants treated with recombinant Tie2–Fc or in control conditions. The z–y–x planes are also depicted to illustrate the positioning of the cells. A scheme of explant preparation is also provided in Extended Data Fig. 2e. g, Quantification of the number of OpCs in the MZ (Olig2+) of Tie2–Fc-treated explants, normalized to vehicle-treated explants. Graph shows mean ± s.e.m. (control n = 13, Tie2–Fc n = 17; 5 independent litters). Unpaired two-sided t-test, *P < 0.0248.

Journal: Nature neuroscience

Article Title: Oligodendrocyte precursor cell specification is regulated by bidirectional neural progenitor–endothelial cell crosstalk

doi: 10.1038/s41593-020-00788-z

Figure Lengend Snippet: a, Ang1 ISH in transverse sections from E10.5 to E13.5 mouse embryos at brachial level (top). Combined Ang1 ISH (pseudo-colored in red) with Olig2 immunofluorescence (bottom). Arrowheads point to Ang1+ co-localization with Olig2+ pMN-NpCs. Scale bar, 50 μm (bottom) and 100 μm (top). b, plot showing that Ang1 expression (% of the area of Ang1+ signal in ventral NpC domains; left y axis) sequentially overlaps with OpC specification (OpC counts; right y axis) at the pMN between E9.5 and E13.5 (see Methods for more details). Graph shows mean ± s.e.m. (Olig2 counts: E9.5: n = 5 images from one embryo, E10.5: n = 20 images from three independent embryos, E11.5: n = 16 images from two independent embryos, E12.5: n = 24 images from four independent embryos, E13.5: n = 15 images from four independent embryos; Ang1 expression: E9.5: n = 6, E10.5: n = 8, E11.5: n = 7, E12.5: n = 14, E13.5: n = 7 images from two independent embryos). c, d, Relative Ptch (c) and Ang1 (d) expression in NSps derived from E11.5 SC progenitors stimulated with recombinant Shh (500 ng ml−1) and/or Shh inhibitor Sant1 (150 nM). Graph shows mean ± s.e.m. (n = 5 independent experiments; one-way ANOVA. Ptch: Control versus Shh *P = 0.0391; Shh versus Shh+Sant1 *P = 0.0142. Ang1: Control versus Shh *P = 0.0375; Shh versus Shh+Sant1 *P = 0.0242). e, Whole-mount SC explants immunostained for Olig2 and Sox10 to detect OpCs (arrowheads) in the VZ and MZ after culturing in control (vehicle) or recombinant Tie2–Fc (2 μg ml−1) in the y–x plane of Z-stack images. On the right side of each image, an orthogonal projection is shown for the z–y plane of a Z-stack. Scale bar, 100 μm. Open arrowheads indicate OpCs in the pMN (VZ) and filled arrowheads in the MZ. f, Scheme summarizing the phenotype of explants treated with recombinant Tie2–Fc or in control conditions. The z–y–x planes are also depicted to illustrate the positioning of the cells. A scheme of explant preparation is also provided in Extended Data Fig. 2e. g, Quantification of the number of OpCs in the MZ (Olig2+) of Tie2–Fc-treated explants, normalized to vehicle-treated explants. Graph shows mean ± s.e.m. (control n = 13, Tie2–Fc n = 17; 5 independent litters). Unpaired two-sided t-test, *P < 0.0248.

Article Snippet: Secondary NSPs (NSP-2) that grew in suspension were stimulated with recombinant Shh 500 ng ml −1 (923-AN, R&D Systems) and/or Sant1 150 nM (559303, Calbiochem) and lysed for RNA extraction.

Techniques: Immunofluorescence, Expressing, Derivative Assay, Recombinant, Control

a, Tgfβ1 ISH in E12.5 wild-type embryo and stained for blood vessels (IsoB4+) (arrowheads show localization of Tgfβ1 mRNA in the vasculature). Scale bar, 100 μm. b,c, Relative expression of Tgfβ1 transcript of HBMECs (b) and isolated E12.5 wild-type SC ECs (c) upon control or recombinant Ang1 stimulation. Graphs show mean ± s.e.m. (b) n = 5 independent experiments, unpaired two-sided t-test *P = 0.0146; (c) n = 6 independent experiments, unpaired two-sided t-test *P = 0.0479). d, Relative expression of Tgfβ1 transcript in HBMECs treated with vehicle (DMSO), recombinant Ang1 and/or Akt/PI3K inhibitor Ly294002. Graph shows mean ± s.e.m. (n = 5 independent experiments). One-way ANOVA, Control versus +Ang1 **P = 0.0077; +Ang1 versus +Ly294002 **P = 0.0012; +Ang1 versus +Ang1+Ly294002 ****P < 0.0001. e,f, Relative expression of Tgfβ1 mRNA in CD31+ ECs sorted from Ang1 fl/fl and Ang1 fl/flNestin:Cre E12.5 SCs (e) and Tie2 fl/fl and Tie2 fl/flPdgfb:CreERT2 SCs (f). Graphs show mean ± s.e.m. (Ang1 fl/fl n = 17, Ang1 fl/flNestin:Cre n = 15, from four independent litters, unpaired two-sided t-test, *P = 0.0495; Tie2 fl/fl n = 11, Tie2 fl/fl Pdgfb: CreERT2 n = 11, from three independent litters, unpaired two-sided t-test, **P = 0.0033). g, Co-immunostaining for phospho-SMAD3 (p-SMAD3) and Olig2 at the pMN level of E12.5 Ang1 fl/fl and Ang1 fl/flNestin:Cre embryo transverse section. Arrowheads show Olig2+pSMAD3+ double-positive cells. Scale bar, 25 μm. h, Quantification of p-SMAD3 fluorescence intensity per Olig2+ cell counts in the pMN of Ang1 fl/fl and Ang1 fl/flNestin:Cre normalized to control littermates. Graph shows mean ± s.e.m. (Ang1 fl/fl n = 10, Ang1 fl/flNestin:Cre n = 10, from three independent litters). Unpaired two-sided t-test, *P = 0.040. AU, arbitrary units. i, Quantification of the number of OPCs in the MZ (Olig2+) from wild-type E11.5 explants treated with α-TGFβ1 blocking antibody (α-TGFβ1 Ab) or vehicle (IgY control) (5 μg ml−1) for 24 h; values are normalized to control-treated explants. Graph shows mean ± s.e.m. (IgY control n = 12, α-TGFβ1 Ab n = 12, from three independent litters). Unpaired two-sided t-test, **P = 0.0088. j, Quantification of OPC number in the MZ (Olig2+) of Ang1 fl/fl and Ang1 fl/flNestin:Cre SC explants after culturing with control (vehicle) or recombinant TGFβ1 50 ng ml−1, normalized to control (vehicle of Ang1 fl/fl). Graph shows mean ± s.e.m. (Ang1 fl/fl n = 10 vehicle and +TGFβ1 n = 11; Ang1 fl/flNestin:Cre n = 18 vehicle and +TGFβ1 n = 10, from seven independent litters). Two-way ANOVA; Control Ang1 fl/fl versus Control Ang1 fl/flNestin:Cre *P = 0.0449; Control Ang1 fl/flNestin:Cre versus +TGFβ1 Ang1 fl/flNestin:Cre **P = 0.0029. k, Quantification of the number of OPCs in the MZ (Olig2+) of Tie2 fl/fl and Tie2 fl/flPdgfb:CreERT2 SC explants after culturing with control (vehicle) or recombinant TGFβ1 50 ng ml−1, normalized to control (vehicle of Tie2 fl/fl). Graph shows mean ± s.e.m. (Tie2 fl/fl n = 15 vehicle and +TGFβ1 n = 5; Tie2 fl/fl Pdgfb:CreERT2 n = 11 vehicle and +TGFβ1 n = 10, from five independent litters). Two-way ANOVA; Control Tie2 fl/fl versus Control Tie2 fl/fl Pdgfb:CreERT2 *P = 0.0278; Control Tie2 fl/fl Pdgfb:CreERT2 versus +TGFβ1 Tie2 fl/fl Pdgfb:CreERT2 ***P = 0.0002.

Journal: Nature neuroscience

Article Title: Oligodendrocyte precursor cell specification is regulated by bidirectional neural progenitor–endothelial cell crosstalk

doi: 10.1038/s41593-020-00788-z

Figure Lengend Snippet: a, Tgfβ1 ISH in E12.5 wild-type embryo and stained for blood vessels (IsoB4+) (arrowheads show localization of Tgfβ1 mRNA in the vasculature). Scale bar, 100 μm. b,c, Relative expression of Tgfβ1 transcript of HBMECs (b) and isolated E12.5 wild-type SC ECs (c) upon control or recombinant Ang1 stimulation. Graphs show mean ± s.e.m. (b) n = 5 independent experiments, unpaired two-sided t-test *P = 0.0146; (c) n = 6 independent experiments, unpaired two-sided t-test *P = 0.0479). d, Relative expression of Tgfβ1 transcript in HBMECs treated with vehicle (DMSO), recombinant Ang1 and/or Akt/PI3K inhibitor Ly294002. Graph shows mean ± s.e.m. (n = 5 independent experiments). One-way ANOVA, Control versus +Ang1 **P = 0.0077; +Ang1 versus +Ly294002 **P = 0.0012; +Ang1 versus +Ang1+Ly294002 ****P < 0.0001. e,f, Relative expression of Tgfβ1 mRNA in CD31+ ECs sorted from Ang1 fl/fl and Ang1 fl/flNestin:Cre E12.5 SCs (e) and Tie2 fl/fl and Tie2 fl/flPdgfb:CreERT2 SCs (f). Graphs show mean ± s.e.m. (Ang1 fl/fl n = 17, Ang1 fl/flNestin:Cre n = 15, from four independent litters, unpaired two-sided t-test, *P = 0.0495; Tie2 fl/fl n = 11, Tie2 fl/fl Pdgfb: CreERT2 n = 11, from three independent litters, unpaired two-sided t-test, **P = 0.0033). g, Co-immunostaining for phospho-SMAD3 (p-SMAD3) and Olig2 at the pMN level of E12.5 Ang1 fl/fl and Ang1 fl/flNestin:Cre embryo transverse section. Arrowheads show Olig2+pSMAD3+ double-positive cells. Scale bar, 25 μm. h, Quantification of p-SMAD3 fluorescence intensity per Olig2+ cell counts in the pMN of Ang1 fl/fl and Ang1 fl/flNestin:Cre normalized to control littermates. Graph shows mean ± s.e.m. (Ang1 fl/fl n = 10, Ang1 fl/flNestin:Cre n = 10, from three independent litters). Unpaired two-sided t-test, *P = 0.040. AU, arbitrary units. i, Quantification of the number of OPCs in the MZ (Olig2+) from wild-type E11.5 explants treated with α-TGFβ1 blocking antibody (α-TGFβ1 Ab) or vehicle (IgY control) (5 μg ml−1) for 24 h; values are normalized to control-treated explants. Graph shows mean ± s.e.m. (IgY control n = 12, α-TGFβ1 Ab n = 12, from three independent litters). Unpaired two-sided t-test, **P = 0.0088. j, Quantification of OPC number in the MZ (Olig2+) of Ang1 fl/fl and Ang1 fl/flNestin:Cre SC explants after culturing with control (vehicle) or recombinant TGFβ1 50 ng ml−1, normalized to control (vehicle of Ang1 fl/fl). Graph shows mean ± s.e.m. (Ang1 fl/fl n = 10 vehicle and +TGFβ1 n = 11; Ang1 fl/flNestin:Cre n = 18 vehicle and +TGFβ1 n = 10, from seven independent litters). Two-way ANOVA; Control Ang1 fl/fl versus Control Ang1 fl/flNestin:Cre *P = 0.0449; Control Ang1 fl/flNestin:Cre versus +TGFβ1 Ang1 fl/flNestin:Cre **P = 0.0029. k, Quantification of the number of OPCs in the MZ (Olig2+) of Tie2 fl/fl and Tie2 fl/flPdgfb:CreERT2 SC explants after culturing with control (vehicle) or recombinant TGFβ1 50 ng ml−1, normalized to control (vehicle of Tie2 fl/fl). Graph shows mean ± s.e.m. (Tie2 fl/fl n = 15 vehicle and +TGFβ1 n = 5; Tie2 fl/fl Pdgfb:CreERT2 n = 11 vehicle and +TGFβ1 n = 10, from five independent litters). Two-way ANOVA; Control Tie2 fl/fl versus Control Tie2 fl/fl Pdgfb:CreERT2 *P = 0.0278; Control Tie2 fl/fl Pdgfb:CreERT2 versus +TGFβ1 Tie2 fl/fl Pdgfb:CreERT2 ***P = 0.0002.

Article Snippet: Secondary NSPs (NSP-2) that grew in suspension were stimulated with recombinant Shh 500 ng ml −1 (923-AN, R&D Systems) and/or Sant1 150 nM (559303, Calbiochem) and lysed for RNA extraction.

Techniques: Staining, Expressing, Isolation, Control, Recombinant, Immunostaining, Fluorescence, Blocking Assay

a, 100-μm transverse sections of Ang1 fl/fl and Ang1 fl/flNestin:Cre SC explants immunostained for Olig2 and Sox10 to detect OPCs in the VZ (open arrowheads) and MZ (filled arrowheads) after culturing with control (vehicle) or recombinant Ang1 300 ng ml−1 for 24 h. Scale bar, 100 μm. b, Quantification of the number of OPCs in the MZ (Olig2+), normalized to control (Ang1 fl/fl) untreated explants. Mean ± s.e.m. (Ang1 fl/fl n = 15 vehicle and n = 14 +Ang1; Ang1 fl/flNestin:Cre n = 15 vehicle and n = 14 +Ang1, from six independent litters). Two-way ANOVA (Ang1 fl/fl versus Ang1 fl/flNestin:Cre (vehicle) *P = 0.0164; Ang1 fl/flNestin:Cre vehicle versus Ang1 fl/flNestin:Cre +Ang1 **P = 0.0015). c, 100-μm transverse sections of Ang1 fl/fl and Ang1 fl/flOlig2:Cre SC explants immunostained for Olig2 and Sox10 to detect OPCs in the VZ (open arrowheads) and MZ (filled arrowheads) after culturing with control (vehicle) or recombinant Ang1 300 ng ml−1 for 24 h. Scale bar, 100 μm. d, Quantification of the number of OPCs in the MZ (Olig2+), normalized to control (Ang1 fl/fl) untreated explants. Graph shows mean ± s.e.m. (Ang1 fl/fl n = 7 vehicle and n = 4 +Ang1, Ang1 fl/flOlig2:Cre n = 3 vehicle and n = 4 +Ang1, from four independent litters). Two-way ANOVA (Control Ang1 fl/fl versus Control Ang1 fl/flOlig2:Cre **P = 0.0093; Control Ang1 fl/flOlig2:Cre versus +Ang1 Ang1 fl/flOlig2:Cre **P = 0.0074).

Journal: Nature neuroscience

Article Title: Oligodendrocyte precursor cell specification is regulated by bidirectional neural progenitor–endothelial cell crosstalk

doi: 10.1038/s41593-020-00788-z

Figure Lengend Snippet: a, 100-μm transverse sections of Ang1 fl/fl and Ang1 fl/flNestin:Cre SC explants immunostained for Olig2 and Sox10 to detect OPCs in the VZ (open arrowheads) and MZ (filled arrowheads) after culturing with control (vehicle) or recombinant Ang1 300 ng ml−1 for 24 h. Scale bar, 100 μm. b, Quantification of the number of OPCs in the MZ (Olig2+), normalized to control (Ang1 fl/fl) untreated explants. Mean ± s.e.m. (Ang1 fl/fl n = 15 vehicle and n = 14 +Ang1; Ang1 fl/flNestin:Cre n = 15 vehicle and n = 14 +Ang1, from six independent litters). Two-way ANOVA (Ang1 fl/fl versus Ang1 fl/flNestin:Cre (vehicle) *P = 0.0164; Ang1 fl/flNestin:Cre vehicle versus Ang1 fl/flNestin:Cre +Ang1 **P = 0.0015). c, 100-μm transverse sections of Ang1 fl/fl and Ang1 fl/flOlig2:Cre SC explants immunostained for Olig2 and Sox10 to detect OPCs in the VZ (open arrowheads) and MZ (filled arrowheads) after culturing with control (vehicle) or recombinant Ang1 300 ng ml−1 for 24 h. Scale bar, 100 μm. d, Quantification of the number of OPCs in the MZ (Olig2+), normalized to control (Ang1 fl/fl) untreated explants. Graph shows mean ± s.e.m. (Ang1 fl/fl n = 7 vehicle and n = 4 +Ang1, Ang1 fl/flOlig2:Cre n = 3 vehicle and n = 4 +Ang1, from four independent litters). Two-way ANOVA (Control Ang1 fl/fl versus Control Ang1 fl/flOlig2:Cre **P = 0.0093; Control Ang1 fl/flOlig2:Cre versus +Ang1 Ang1 fl/flOlig2:Cre **P = 0.0074).

Article Snippet: Secondary NSPs (NSP-2) that grew in suspension were stimulated with recombinant Shh 500 ng ml −1 (923-AN, R&D Systems) and/or Sant1 150 nM (559303, Calbiochem) and lysed for RNA extraction.

Techniques: Control, Recombinant

Parkin and SLP-2 interact in human control fibroblasts and hiPSC-derived neurons. A Representative PLA staining for fibroblasts of a control individual under normal culture conditions and after CCCP treatment (3 h, 10 µM). The PLA signal is visualized in red, DAPI-stained nuclei are shown in blue. Scale bar: 10 µm. Quantification of PLA dots per cell shows a significantly higher PLA signal after CCCP exposure, indicating an increased interaction between the two proteins. Statistical differences were calculated by unpaired Mann Whitney U test ****p ≤ 0.0001. B hiPSC-derived neurons of a control individual were processed using PLA under normal culture conditions and after CCCP treatment (3 h, 10 µM). Scale bar: 10 µm. Quantification of PLA dots per cell shows a significantly higher PLA signal after CCCP exposure. Statistical differences were calculated by unpaired Mann Whitney U test. **p ≤ 0.005. The specificity of the PLA interaction results was confirmed by performing the experiments with only one of the two primary antibodies. C Far-Western blot analysis using recombinant SLP-2 (aa 41–356) and Parkin proteins (aa 1–465 and aa 138–465) shows the direct binding of the two proteins. D Quantification of PLA dots per cell in control and PRKN mutant fibroblast lines under normal culture conditions and after CCCP treatment (3 h, 10 µM) using an antibody directed against the Parkin N-terminus and an anti-SLP-2 antibody. Only in the control lines, CCCP treatment resulted in a significantly increased interaction. Statistical differences were calculated by one-way ANOVA followed by Tukey’s post hoc test to correct for multiple comparisons. * ≤ 0.05, **p ≤ 0.01, ****p ≤ 0.0001

Journal: Journal of Translational Medicine

Article Title: Intracellular delivery of Parkin-RING0-based fragments corrects Parkin-induced mitochondrial dysfunction through interaction with SLP-2

doi: 10.1186/s12967-024-04850-3

Figure Lengend Snippet: Parkin and SLP-2 interact in human control fibroblasts and hiPSC-derived neurons. A Representative PLA staining for fibroblasts of a control individual under normal culture conditions and after CCCP treatment (3 h, 10 µM). The PLA signal is visualized in red, DAPI-stained nuclei are shown in blue. Scale bar: 10 µm. Quantification of PLA dots per cell shows a significantly higher PLA signal after CCCP exposure, indicating an increased interaction between the two proteins. Statistical differences were calculated by unpaired Mann Whitney U test ****p ≤ 0.0001. B hiPSC-derived neurons of a control individual were processed using PLA under normal culture conditions and after CCCP treatment (3 h, 10 µM). Scale bar: 10 µm. Quantification of PLA dots per cell shows a significantly higher PLA signal after CCCP exposure. Statistical differences were calculated by unpaired Mann Whitney U test. **p ≤ 0.005. The specificity of the PLA interaction results was confirmed by performing the experiments with only one of the two primary antibodies. C Far-Western blot analysis using recombinant SLP-2 (aa 41–356) and Parkin proteins (aa 1–465 and aa 138–465) shows the direct binding of the two proteins. D Quantification of PLA dots per cell in control and PRKN mutant fibroblast lines under normal culture conditions and after CCCP treatment (3 h, 10 µM) using an antibody directed against the Parkin N-terminus and an anti-SLP-2 antibody. Only in the control lines, CCCP treatment resulted in a significantly increased interaction. Statistical differences were calculated by one-way ANOVA followed by Tukey’s post hoc test to correct for multiple comparisons. * ≤ 0.05, **p ≤ 0.01, ****p ≤ 0.0001

Article Snippet: On days 1 to 5, KSR medium was added to the cells in the presence of SB, LDN, recombinant Human Sonic Hedgehog (SHH, R&D System), recombinant Human FGF-8a (FGF8, R&D System), and Purmorphamine (Pu, StemMACS).

Techniques: Control, Derivative Assay, Staining, MANN-WHITNEY, Far Western Blot, Recombinant, Binding Assay, Mutagenesis