Chromatin Immunoprecipitation:Article Title: Control of neuronal excitation–inhibition balance by BMP–SMAD1 signalling
Article Snippet: Clozapine N-oxide (CNO) (5 mg kg −1 , Sigma Aldrich, C0832) and doxycycline (50 mg kg −1 , Thermo Fisher Scientific, BP26531, suspended in 0.9% NaCl to 5 mg ml −1 concentration) were administered by intraperitoneal injection. .. Primary antibodies were: monoclonal mouse anti-synaptotagmin-2 (Zebrafish International Resource Center, ZNP-1), rabbit anti-SMAD1 (Cell Signaling 6944, 1:100 for ChIP and 1:1,000 for western blot), H3K27ac (Abcam 4729, 1:1,000), rabbit anti-SMAD5 (Cell Signaling, 12534, 1:100 for ChIP and 1:1,000 for western blot), anti-phospho-SMAD1/5/9 (Cell Signaling 13820, 1:1,000), mouse anti-BMPR2 (BD Pharmingen, 612292, 1:1,000), rabbit anti-calnexin (StressGen, SPA-865, 1:2,000), mouse anti-MAP2 (Synaptic Systems, 188011, 1:1,000), mouse anti-CAMKII alpha (Thermo Fisher Scientific, MA1-048, 1:800), rat anti-GAPDH (Biolegend, 607902, 1:10,000), rabbit anti-NeuN (Abcam, ab177487, 1:500), mouse anti-GAD67 (Millipore MAB5406, 1:500), rabbit anti-vGLUT1 (Synaptic Systems 135303, 1:5,000), biotinylated WFA (Vector Laboratories B-1355-2, 1:500), rabbit anti-HA (Cell Signaling 3724, 1:1,000), mouse anti-GFP antibody (Santa Cruz, sc-9996, 1:1,000) and goat anti-parvalbumin antibody (Swant PVG213, 1:5,000). .. Secondary antibodies were: HRP goat anti-rabbit (Jackson 111-035-003, 1:10,000), HRP goat anti-rat (Jackson 112-035-143, 1:10,000), HRP goat anti-mouse (Jackson 115-035-149, 1:10,000), Alexa405 goat anti-rabbit (Thermo Fisher Scientific A-31556, 1:500), Alexa488 donkey anti-rabbit (Thermo Fisher Scientific R37118, 1:1,000), Alexa647 donkey anti-mouse (Jackson 715-605-151, 1:1,000), Alexa647 streptavidin (Thermo Fisher Scientific, S32357, 1:1,000), Cy2 Streptavidin (Jackson 016-220-084, 1:1,000), Cy3 donkey anti-mouse (Jackson 715-165-151, 1:500), Cy3 donkey anti-rabbit (Jackson 711-165-152, 1:500), Cy5 donkey anti-goat (Jackson 705-175-147, 1:500), Cy5 donkey anti-rabbit (Jackson 711-175-152, 1:500) and Cy5 donkey anti-mouse (Jackson 715-175-511, 1:500).
Article Title: Control of neuronal excitation-inhibition balance by BMP-SMAD1 signalling.
Article Snippet: Clozapine N-oxide (CNO) (5 mg kg−1, Sigma Aldrich, C0832) and doxycycline (50 mg kg−1, Thermo Fisher Scientific, BP26531, suspended in 0.9% NaCl to 5 mg ml−1 concentration) were administered by intraperitoneal injection. .. Primary antibodies were: monoclonal mouse anti-synaptotagmin-2 (Zebrafish International Resource Center, ZNP-1), rabbit anti-SMAD1 (Cell Signaling 6944, 1:100 for ChIP and 1:1,000 for western blot), H3K27ac (Abcam 4729, 1:1,000), rabbit anti-SMAD5 (Cell Signaling, 12534, 1:100 for ChIP and 1:1,000 for western blot), anti-phospho-SMAD1/5/9 (Cell Signaling 13820, 1:1,000), mouse anti-BMPR2 (BD Pharmingen, 612292, 1:1,000), rabbit anti-calnexin (StressGen, SPA-865, 1:2,000), mouse anti-MAP2 (Synaptic Systems, 188011, 1:1,000), mouse anti-CAMKII alpha (Thermo Fisher Scientific, MA1-048, 1:800), rat anti-GAPDH (Biolegend, 607902, 1:10,000), rabbit anti-NeuN (Abcam, ab177487, 1:500), mouse anti-GAD67 (Millipore MAB5406, 1:500), rabbit anti-vGLUT1 (Synaptic Systems 135303, 1:5,000), biotinylated WFA (Vector Laboratories B-1355-2, 1:500), rabbit anti-HA (Cell Signaling 3724, 1:1,000), mouse anti-GFP antibody (Santa Cruz, sc-9996, 1:1,000) and goat anti-parvalbumin antibody (Swant PVG213, 1:5,000). .. Secondary antibodies were: HRP goat anti-rabbit ( Jackson 111-035-003, 1:10,000), HRP goat anti-rat ( Jackson 112-035-143, 1:10,000), HRP goat anti-mouse ( Jackson 115-035-149, 1:10,000), Alexa405 goat anti-rabbit (Thermo Fisher Scientific A-31556, 1:500), Alexa488 donkey anti-rabbit (Thermo Fisher Scientific R37118, 1:1,000), Alexa647 donkey anti-mouse ( Jackson 715-605-151, 1:1,000), Alexa647 streptavidin (Thermo Fisher Scientific, S32357, 1:1,000), Cy2 Streptavidin ( Jackson 016-220-084, 1:1,000), Cy3 donkey anti-mouse ( Jackson 715-165-151, 1:500), Cy3 donkey anti-rabbit ( Jackson 711-165- 152, 1:500), Cy5 donkey anti-goat ( Jackson 705-175-147, 1:500), Cy5 donkey anti-rabbit ( Jackson 711-175-152, 1:500) and Cy5 donkey anti-mouse ( Jackson 715-175-511, 1:500).
Western Blot:Article Title: Control of neuronal excitation–inhibition balance by BMP–SMAD1 signalling
Article Snippet: Clozapine N-oxide (CNO) (5 mg kg −1 , Sigma Aldrich, C0832) and doxycycline (50 mg kg −1 , Thermo Fisher Scientific, BP26531, suspended in 0.9% NaCl to 5 mg ml −1 concentration) were administered by intraperitoneal injection. .. Primary antibodies were: monoclonal mouse anti-synaptotagmin-2 (Zebrafish International Resource Center, ZNP-1), rabbit anti-SMAD1 (Cell Signaling 6944, 1:100 for ChIP and 1:1,000 for western blot), H3K27ac (Abcam 4729, 1:1,000), rabbit anti-SMAD5 (Cell Signaling, 12534, 1:100 for ChIP and 1:1,000 for western blot), anti-phospho-SMAD1/5/9 (Cell Signaling 13820, 1:1,000), mouse anti-BMPR2 (BD Pharmingen, 612292, 1:1,000), rabbit anti-calnexin (StressGen, SPA-865, 1:2,000), mouse anti-MAP2 (Synaptic Systems, 188011, 1:1,000), mouse anti-CAMKII alpha (Thermo Fisher Scientific, MA1-048, 1:800), rat anti-GAPDH (Biolegend, 607902, 1:10,000), rabbit anti-NeuN (Abcam, ab177487, 1:500), mouse anti-GAD67 (Millipore MAB5406, 1:500), rabbit anti-vGLUT1 (Synaptic Systems 135303, 1:5,000), biotinylated WFA (Vector Laboratories B-1355-2, 1:500), rabbit anti-HA (Cell Signaling 3724, 1:1,000), mouse anti-GFP antibody (Santa Cruz, sc-9996, 1:1,000) and goat anti-parvalbumin antibody (Swant PVG213, 1:5,000). .. Secondary antibodies were: HRP goat anti-rabbit (Jackson 111-035-003, 1:10,000), HRP goat anti-rat (Jackson 112-035-143, 1:10,000), HRP goat anti-mouse (Jackson 115-035-149, 1:10,000), Alexa405 goat anti-rabbit (Thermo Fisher Scientific A-31556, 1:500), Alexa488 donkey anti-rabbit (Thermo Fisher Scientific R37118, 1:1,000), Alexa647 donkey anti-mouse (Jackson 715-605-151, 1:1,000), Alexa647 streptavidin (Thermo Fisher Scientific, S32357, 1:1,000), Cy2 Streptavidin (Jackson 016-220-084, 1:1,000), Cy3 donkey anti-mouse (Jackson 715-165-151, 1:500), Cy3 donkey anti-rabbit (Jackson 711-165-152, 1:500), Cy5 donkey anti-goat (Jackson 705-175-147, 1:500), Cy5 donkey anti-rabbit (Jackson 711-175-152, 1:500) and Cy5 donkey anti-mouse (Jackson 715-175-511, 1:500).
Article Title: R-Spondin 2 governs Xenopus left-right body axis formation by establishing an FGF signaling gradient
Article Snippet: .. Primary antibodies used for western blotting: Rabbit anti-Phospho-ERK1/2 (Cell Signaling Technology 9101 S); Rabbit anti-ERK1/2 (Sigma M5670); Mouse anti-beta-Catenin (BD 610154); Rabbit anti-LRP6 (Cell Signaling Technology 2560); Rabbit anti-GAPDH (Cell Signaling Technology 2118 S); Goat anti-RSPO2 (R and D systems AF3266); Rat anti-HA (Roche 11867423001); Rabbit anti-Phospho-Smad1 (Cell Signaling Technology 9516); Rabbit anti-Smad1(Cell Signaling Technology 9743 S). .. Secondary antibodies used for western blotting: Goat anti-mouse IgG (H + L) HRP (Jackson ImmunoResearch 115-035-146); Goat anti-rabbit IgG (H + L) HRP (Jackson ImmunoResearch 111-035-144); Mouse anti-goat IgG (H + L) HRP (Jackson ImmunoResearch 205-035-108).
Article Title: R-Spondin 2 governs Xenopus left-right body axis formation by establishing an FGF signaling gradient
Article Snippet: .. Primary antibodies used for Xenopus laevis western blotting: Rabbit anti-Phospho-ERK1/2 (Cell Signaling Technology 9101 S); Rabbit anti-ERK1/2 (Sigma M5670); Rabbit anti-Phospho-Smad1 (Cell Signaling Technology 9516); Rabbit anti-Smad1(Cell Signaling Technology 9743 S); Rabbit anti-GAPDH (Cell Signaling Technology 2118 S); Mouse anti-active-beta-catenin (Millipore 05-665); Rabbit anti-ERK1/2 (GeneTex GTX134462). .. Secondary antibodies used for Xenopus laevis western blotting: Goat anti-mouse IgG (H + L) HRP (Jackson ImmunoResearch 115-035-146); Goat anti-rabbit IgG (H + L) HRP (Jackson ImmunoResearch 111-035-144).
Article Title: Allosteric AKT inhibitors for use in the treatment of Hereditary Hemorrhagic Telangiectasia
Article Snippet: Complete high-resolution 3-dimensional rendering of whole-mount retinas was acquired using Laser Scanning Microscope SP5 or SP8 (Leica). .. Western Blot and Quantification Samples were boiled for 10 min and proteins were separated on 10% acrylamide gel and transferred onto nitrocellulose membrane before blocking with 5% BSA or 5% powdered milk/Tris-buffered saline/Tween and incubated with the following primary antibodies: rabbit anti-Phospho-Akt1 (Ser473) (44-621G, Invitrogen), rabbit anti-Akt1 (2938S, Cell Signaling), rabbit anti-PhosphoSmad1/5 (Ser463/465) (9516S, Cell Signaling), rabbit anti-Smad1 (6944S, Cell Signaling), mouse anti-Phospho-p44/42 MAPK (Thr202/Tyr204) (E10) (9106S, Cell signaling), rabbit p44/42 MAPK (Erk1/2) (9102S, Cell Signaling), goat anti-Endoglin (AF1097, R&D systems) and mouse anti-β-actin (A5441, Sigma-Aldrich). .. Detection was achieved using HRP anti-rabbit IgG or HRP anti-mouse IgG (W4011 or W4021 respectively, Promega) or HRP anti-goat IgG (HAF017, R&D systems) followed by scanning on BIO-RAD ChemiDoc imager.
Article Title: Control of neuronal excitation-inhibition balance by BMP-SMAD1 signalling.
Article Snippet: Clozapine N-oxide (CNO) (5 mg kg−1, Sigma Aldrich, C0832) and doxycycline (50 mg kg−1, Thermo Fisher Scientific, BP26531, suspended in 0.9% NaCl to 5 mg ml−1 concentration) were administered by intraperitoneal injection. .. Primary antibodies were: monoclonal mouse anti-synaptotagmin-2 (Zebrafish International Resource Center, ZNP-1), rabbit anti-SMAD1 (Cell Signaling 6944, 1:100 for ChIP and 1:1,000 for western blot), H3K27ac (Abcam 4729, 1:1,000), rabbit anti-SMAD5 (Cell Signaling, 12534, 1:100 for ChIP and 1:1,000 for western blot), anti-phospho-SMAD1/5/9 (Cell Signaling 13820, 1:1,000), mouse anti-BMPR2 (BD Pharmingen, 612292, 1:1,000), rabbit anti-calnexin (StressGen, SPA-865, 1:2,000), mouse anti-MAP2 (Synaptic Systems, 188011, 1:1,000), mouse anti-CAMKII alpha (Thermo Fisher Scientific, MA1-048, 1:800), rat anti-GAPDH (Biolegend, 607902, 1:10,000), rabbit anti-NeuN (Abcam, ab177487, 1:500), mouse anti-GAD67 (Millipore MAB5406, 1:500), rabbit anti-vGLUT1 (Synaptic Systems 135303, 1:5,000), biotinylated WFA (Vector Laboratories B-1355-2, 1:500), rabbit anti-HA (Cell Signaling 3724, 1:1,000), mouse anti-GFP antibody (Santa Cruz, sc-9996, 1:1,000) and goat anti-parvalbumin antibody (Swant PVG213, 1:5,000). .. Secondary antibodies were: HRP goat anti-rabbit ( Jackson 111-035-003, 1:10,000), HRP goat anti-rat ( Jackson 112-035-143, 1:10,000), HRP goat anti-mouse ( Jackson 115-035-149, 1:10,000), Alexa405 goat anti-rabbit (Thermo Fisher Scientific A-31556, 1:500), Alexa488 donkey anti-rabbit (Thermo Fisher Scientific R37118, 1:1,000), Alexa647 donkey anti-mouse ( Jackson 715-605-151, 1:1,000), Alexa647 streptavidin (Thermo Fisher Scientific, S32357, 1:1,000), Cy2 Streptavidin ( Jackson 016-220-084, 1:1,000), Cy3 donkey anti-mouse ( Jackson 715-165-151, 1:500), Cy3 donkey anti-rabbit ( Jackson 711-165- 152, 1:500), Cy5 donkey anti-goat ( Jackson 705-175-147, 1:500), Cy5 donkey anti-rabbit ( Jackson 711-175-152, 1:500) and Cy5 donkey anti-mouse ( Jackson 715-175-511, 1:500).
Acrylamide Gel Assay:Article Title: Allosteric AKT inhibitors for use in the treatment of Hereditary Hemorrhagic Telangiectasia
Article Snippet: Complete high-resolution 3-dimensional rendering of whole-mount retinas was acquired using Laser Scanning Microscope SP5 or SP8 (Leica). .. Western Blot and Quantification Samples were boiled for 10 min and proteins were separated on 10% acrylamide gel and transferred onto nitrocellulose membrane before blocking with 5% BSA or 5% powdered milk/Tris-buffered saline/Tween and incubated with the following primary antibodies: rabbit anti-Phospho-Akt1 (Ser473) (44-621G, Invitrogen), rabbit anti-Akt1 (2938S, Cell Signaling), rabbit anti-PhosphoSmad1/5 (Ser463/465) (9516S, Cell Signaling), rabbit anti-Smad1 (6944S, Cell Signaling), mouse anti-Phospho-p44/42 MAPK (Thr202/Tyr204) (E10) (9106S, Cell signaling), rabbit p44/42 MAPK (Erk1/2) (9102S, Cell Signaling), goat anti-Endoglin (AF1097, R&D systems) and mouse anti-β-actin (A5441, Sigma-Aldrich). .. Detection was achieved using HRP anti-rabbit IgG or HRP anti-mouse IgG (W4011 or W4021 respectively, Promega) or HRP anti-goat IgG (HAF017, R&D systems) followed by scanning on BIO-RAD ChemiDoc imager.
Membrane:Article Title: Allosteric AKT inhibitors for use in the treatment of Hereditary Hemorrhagic Telangiectasia
Article Snippet: Complete high-resolution 3-dimensional rendering of whole-mount retinas was acquired using Laser Scanning Microscope SP5 or SP8 (Leica). .. Western Blot and Quantification Samples were boiled for 10 min and proteins were separated on 10% acrylamide gel and transferred onto nitrocellulose membrane before blocking with 5% BSA or 5% powdered milk/Tris-buffered saline/Tween and incubated with the following primary antibodies: rabbit anti-Phospho-Akt1 (Ser473) (44-621G, Invitrogen), rabbit anti-Akt1 (2938S, Cell Signaling), rabbit anti-PhosphoSmad1/5 (Ser463/465) (9516S, Cell Signaling), rabbit anti-Smad1 (6944S, Cell Signaling), mouse anti-Phospho-p44/42 MAPK (Thr202/Tyr204) (E10) (9106S, Cell signaling), rabbit p44/42 MAPK (Erk1/2) (9102S, Cell Signaling), goat anti-Endoglin (AF1097, R&D systems) and mouse anti-β-actin (A5441, Sigma-Aldrich). .. Detection was achieved using HRP anti-rabbit IgG or HRP anti-mouse IgG (W4011 or W4021 respectively, Promega) or HRP anti-goat IgG (HAF017, R&D systems) followed by scanning on BIO-RAD ChemiDoc imager.
Blocking Assay:Article Title: Allosteric AKT inhibitors for use in the treatment of Hereditary Hemorrhagic Telangiectasia
Article Snippet: Complete high-resolution 3-dimensional rendering of whole-mount retinas was acquired using Laser Scanning Microscope SP5 or SP8 (Leica). .. Western Blot and Quantification Samples were boiled for 10 min and proteins were separated on 10% acrylamide gel and transferred onto nitrocellulose membrane before blocking with 5% BSA or 5% powdered milk/Tris-buffered saline/Tween and incubated with the following primary antibodies: rabbit anti-Phospho-Akt1 (Ser473) (44-621G, Invitrogen), rabbit anti-Akt1 (2938S, Cell Signaling), rabbit anti-PhosphoSmad1/5 (Ser463/465) (9516S, Cell Signaling), rabbit anti-Smad1 (6944S, Cell Signaling), mouse anti-Phospho-p44/42 MAPK (Thr202/Tyr204) (E10) (9106S, Cell signaling), rabbit p44/42 MAPK (Erk1/2) (9102S, Cell Signaling), goat anti-Endoglin (AF1097, R&D systems) and mouse anti-β-actin (A5441, Sigma-Aldrich). .. Detection was achieved using HRP anti-rabbit IgG or HRP anti-mouse IgG (W4011 or W4021 respectively, Promega) or HRP anti-goat IgG (HAF017, R&D systems) followed by scanning on BIO-RAD ChemiDoc imager.
Saline:Article Title: Allosteric AKT inhibitors for use in the treatment of Hereditary Hemorrhagic Telangiectasia
Article Snippet: Complete high-resolution 3-dimensional rendering of whole-mount retinas was acquired using Laser Scanning Microscope SP5 or SP8 (Leica). .. Western Blot and Quantification Samples were boiled for 10 min and proteins were separated on 10% acrylamide gel and transferred onto nitrocellulose membrane before blocking with 5% BSA or 5% powdered milk/Tris-buffered saline/Tween and incubated with the following primary antibodies: rabbit anti-Phospho-Akt1 (Ser473) (44-621G, Invitrogen), rabbit anti-Akt1 (2938S, Cell Signaling), rabbit anti-PhosphoSmad1/5 (Ser463/465) (9516S, Cell Signaling), rabbit anti-Smad1 (6944S, Cell Signaling), mouse anti-Phospho-p44/42 MAPK (Thr202/Tyr204) (E10) (9106S, Cell signaling), rabbit p44/42 MAPK (Erk1/2) (9102S, Cell Signaling), goat anti-Endoglin (AF1097, R&D systems) and mouse anti-β-actin (A5441, Sigma-Aldrich). .. Detection was achieved using HRP anti-rabbit IgG or HRP anti-mouse IgG (W4011 or W4021 respectively, Promega) or HRP anti-goat IgG (HAF017, R&D systems) followed by scanning on BIO-RAD ChemiDoc imager.
Incubation:Article Title: Allosteric AKT inhibitors for use in the treatment of Hereditary Hemorrhagic Telangiectasia
Article Snippet: Complete high-resolution 3-dimensional rendering of whole-mount retinas was acquired using Laser Scanning Microscope SP5 or SP8 (Leica). .. Western Blot and Quantification Samples were boiled for 10 min and proteins were separated on 10% acrylamide gel and transferred onto nitrocellulose membrane before blocking with 5% BSA or 5% powdered milk/Tris-buffered saline/Tween and incubated with the following primary antibodies: rabbit anti-Phospho-Akt1 (Ser473) (44-621G, Invitrogen), rabbit anti-Akt1 (2938S, Cell Signaling), rabbit anti-PhosphoSmad1/5 (Ser463/465) (9516S, Cell Signaling), rabbit anti-Smad1 (6944S, Cell Signaling), mouse anti-Phospho-p44/42 MAPK (Thr202/Tyr204) (E10) (9106S, Cell signaling), rabbit p44/42 MAPK (Erk1/2) (9102S, Cell Signaling), goat anti-Endoglin (AF1097, R&D systems) and mouse anti-β-actin (A5441, Sigma-Aldrich). .. Detection was achieved using HRP anti-rabbit IgG or HRP anti-mouse IgG (W4011 or W4021 respectively, Promega) or HRP anti-goat IgG (HAF017, R&D systems) followed by scanning on BIO-RAD ChemiDoc imager.
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