p4 gel filtration column Search Results


p-4 vario planetary mill  (FRITSCH GmbH)
96
FRITSCH GmbH p-4 vario planetary mill
P 4 Vario Planetary Mill, supplied by FRITSCH GmbH, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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isoform ins  (Echelon Biosciences)
94
Echelon Biosciences isoform ins
Analysis of the signaling pathway leading to Bim increased expression. (a) Western blot for Bim expression in splenic B cells from Itpkb+/+ Eμ-2–22 BCL-2 and Itpkb−/− Eμ-2–22 BCL-2 stimulated with goat F(ab′)2 anti-mouse IgM (10 μg/ml) for different times. β-Actin was used as control. The blot is representative of three independent experiments. (b) Analysis of Rasa3 expression and Erk1/2 and BimEL phosphorylation after splenic B cells activation with goat F(ab′)2 anti-mouse IgM (10 μg/ml) for different times or ionomycin (50 ng/ml) plus phorbol 12-myristate 13-acetate (10 ng/ml) for 5 min (I+P). Erk1/2 and β-actin were used as controls. The blot represents one of three independent experiments. (c) Rasa3–GFP subcellular distribution in Cos-7 cells transfected with Rasa3-GFP alone (Upper) or with Rasa3-GFP and Itpkb (Lower). Pictures of representative cells are presented 0, 3, and 5 min after the addition of 100 μM ATP. The profile of fluorescence intensity from membrane to membrane (as depicted by axis presented in small pictures at t = 0) is given below each cell. The fluorescence intensity is represented as arbitrary units. The ratio (R) between the mean fluorescence intensities of the membranes (A1 and A2) and the mean fluorescence intensity of cytosol (B) is given. The pictures are representative of three independent transfection experiments. (d) Ins(1,3,4,5)P4 decreases membrane localization of Rasa3–GFP in HeLa cells. Rasa3–GFP-transfected HeLa cells were cultured for 30 min with DMSO-containing solvent, <t>2,6-di-O-butyryl-Ins(1,2,4,5)P4/PM</t> (50 μg/ml) or 2,6-di-O-butyryl-Ins(1,3,4,5)P4/AM (50 μg/ml). Pictures of representative cells are presented 0 and 30 min after addition of each compound. Data are representative of three independent experiments.
Isoform Ins, supplied by Echelon Biosciences, 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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bio gel p 4 column  (Bio-Rad)
95
Bio-Rad bio gel p 4 column
Analysis of the signaling pathway leading to Bim increased expression. (a) Western blot for Bim expression in splenic B cells from Itpkb+/+ Eμ-2–22 BCL-2 and Itpkb−/− Eμ-2–22 BCL-2 stimulated with goat F(ab′)2 anti-mouse IgM (10 μg/ml) for different times. β-Actin was used as control. The blot is representative of three independent experiments. (b) Analysis of Rasa3 expression and Erk1/2 and BimEL phosphorylation after splenic B cells activation with goat F(ab′)2 anti-mouse IgM (10 μg/ml) for different times or ionomycin (50 ng/ml) plus phorbol 12-myristate 13-acetate (10 ng/ml) for 5 min (I+P). Erk1/2 and β-actin were used as controls. The blot represents one of three independent experiments. (c) Rasa3–GFP subcellular distribution in Cos-7 cells transfected with Rasa3-GFP alone (Upper) or with Rasa3-GFP and Itpkb (Lower). Pictures of representative cells are presented 0, 3, and 5 min after the addition of 100 μM ATP. The profile of fluorescence intensity from membrane to membrane (as depicted by axis presented in small pictures at t = 0) is given below each cell. The fluorescence intensity is represented as arbitrary units. The ratio (R) between the mean fluorescence intensities of the membranes (A1 and A2) and the mean fluorescence intensity of cytosol (B) is given. The pictures are representative of three independent transfection experiments. (d) Ins(1,3,4,5)P4 decreases membrane localization of Rasa3–GFP in HeLa cells. Rasa3–GFP-transfected HeLa cells were cultured for 30 min with DMSO-containing solvent, <t>2,6-di-O-butyryl-Ins(1,2,4,5)P4/PM</t> (50 μg/ml) or 2,6-di-O-butyryl-Ins(1,3,4,5)P4/AM (50 μg/ml). Pictures of representative cells are presented 0 and 30 min after addition of each compound. Data are representative of three independent experiments.
Bio Gel P 4 Column, supplied by Bio-Rad, 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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gel filtration column  (Bio-Rad)
96
Bio-Rad gel filtration column
Analysis of the signaling pathway leading to Bim increased expression. (a) Western blot for Bim expression in splenic B cells from Itpkb+/+ Eμ-2–22 BCL-2 and Itpkb−/− Eμ-2–22 BCL-2 stimulated with goat F(ab′)2 anti-mouse IgM (10 μg/ml) for different times. β-Actin was used as control. The blot is representative of three independent experiments. (b) Analysis of Rasa3 expression and Erk1/2 and BimEL phosphorylation after splenic B cells activation with goat F(ab′)2 anti-mouse IgM (10 μg/ml) for different times or ionomycin (50 ng/ml) plus phorbol 12-myristate 13-acetate (10 ng/ml) for 5 min (I+P). Erk1/2 and β-actin were used as controls. The blot represents one of three independent experiments. (c) Rasa3–GFP subcellular distribution in Cos-7 cells transfected with Rasa3-GFP alone (Upper) or with Rasa3-GFP and Itpkb (Lower). Pictures of representative cells are presented 0, 3, and 5 min after the addition of 100 μM ATP. The profile of fluorescence intensity from membrane to membrane (as depicted by axis presented in small pictures at t = 0) is given below each cell. The fluorescence intensity is represented as arbitrary units. The ratio (R) between the mean fluorescence intensities of the membranes (A1 and A2) and the mean fluorescence intensity of cytosol (B) is given. The pictures are representative of three independent transfection experiments. (d) Ins(1,3,4,5)P4 decreases membrane localization of Rasa3–GFP in HeLa cells. Rasa3–GFP-transfected HeLa cells were cultured for 30 min with DMSO-containing solvent, <t>2,6-di-O-butyryl-Ins(1,2,4,5)P4/PM</t> (50 μg/ml) or 2,6-di-O-butyryl-Ins(1,3,4,5)P4/AM (50 μg/ml). Pictures of representative cells are presented 0 and 30 min after addition of each compound. Data are representative of three independent experiments.
Gel Filtration Column, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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bio-gel p4 gel filtration  (Oxford Glycosystems)
90
Oxford Glycosystems bio-gel p4 gel filtration
Analysis of the signaling pathway leading to Bim increased expression. (a) Western blot for Bim expression in splenic B cells from Itpkb+/+ Eμ-2–22 BCL-2 and Itpkb−/− Eμ-2–22 BCL-2 stimulated with goat F(ab′)2 anti-mouse IgM (10 μg/ml) for different times. β-Actin was used as control. The blot is representative of three independent experiments. (b) Analysis of Rasa3 expression and Erk1/2 and BimEL phosphorylation after splenic B cells activation with goat F(ab′)2 anti-mouse IgM (10 μg/ml) for different times or ionomycin (50 ng/ml) plus phorbol 12-myristate 13-acetate (10 ng/ml) for 5 min (I+P). Erk1/2 and β-actin were used as controls. The blot represents one of three independent experiments. (c) Rasa3–GFP subcellular distribution in Cos-7 cells transfected with Rasa3-GFP alone (Upper) or with Rasa3-GFP and Itpkb (Lower). Pictures of representative cells are presented 0, 3, and 5 min after the addition of 100 μM ATP. The profile of fluorescence intensity from membrane to membrane (as depicted by axis presented in small pictures at t = 0) is given below each cell. The fluorescence intensity is represented as arbitrary units. The ratio (R) between the mean fluorescence intensities of the membranes (A1 and A2) and the mean fluorescence intensity of cytosol (B) is given. The pictures are representative of three independent transfection experiments. (d) Ins(1,3,4,5)P4 decreases membrane localization of Rasa3–GFP in HeLa cells. Rasa3–GFP-transfected HeLa cells were cultured for 30 min with DMSO-containing solvent, <t>2,6-di-O-butyryl-Ins(1,2,4,5)P4/PM</t> (50 μg/ml) or 2,6-di-O-butyryl-Ins(1,3,4,5)P4/AM (50 μg/ml). Pictures of representative cells are presented 0 and 30 min after addition of each compound. Data are representative of three independent experiments.
Bio Gel P4 Gel Filtration, supplied by Oxford Glycosystems, 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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single bio gel p 4 column  (Bio-Rad)
93
Bio-Rad single bio gel p 4 column
Analysis of the signaling pathway leading to Bim increased expression. (a) Western blot for Bim expression in splenic B cells from Itpkb+/+ Eμ-2–22 BCL-2 and Itpkb−/− Eμ-2–22 BCL-2 stimulated with goat F(ab′)2 anti-mouse IgM (10 μg/ml) for different times. β-Actin was used as control. The blot is representative of three independent experiments. (b) Analysis of Rasa3 expression and Erk1/2 and BimEL phosphorylation after splenic B cells activation with goat F(ab′)2 anti-mouse IgM (10 μg/ml) for different times or ionomycin (50 ng/ml) plus phorbol 12-myristate 13-acetate (10 ng/ml) for 5 min (I+P). Erk1/2 and β-actin were used as controls. The blot represents one of three independent experiments. (c) Rasa3–GFP subcellular distribution in Cos-7 cells transfected with Rasa3-GFP alone (Upper) or with Rasa3-GFP and Itpkb (Lower). Pictures of representative cells are presented 0, 3, and 5 min after the addition of 100 μM ATP. The profile of fluorescence intensity from membrane to membrane (as depicted by axis presented in small pictures at t = 0) is given below each cell. The fluorescence intensity is represented as arbitrary units. The ratio (R) between the mean fluorescence intensities of the membranes (A1 and A2) and the mean fluorescence intensity of cytosol (B) is given. The pictures are representative of three independent transfection experiments. (d) Ins(1,3,4,5)P4 decreases membrane localization of Rasa3–GFP in HeLa cells. Rasa3–GFP-transfected HeLa cells were cultured for 30 min with DMSO-containing solvent, <t>2,6-di-O-butyryl-Ins(1,2,4,5)P4/PM</t> (50 μg/ml) or 2,6-di-O-butyryl-Ins(1,3,4,5)P4/AM (50 μg/ml). Pictures of representative cells are presented 0 and 30 min after addition of each compound. Data are representative of three independent experiments.
Single Bio Gel P 4 Column, supplied by Bio-Rad, 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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bio gel p 4 columns  (Bio-Rad)
93
Bio-Rad bio gel p 4 columns
Analysis of the signaling pathway leading to Bim increased expression. (a) Western blot for Bim expression in splenic B cells from Itpkb+/+ Eμ-2–22 BCL-2 and Itpkb−/− Eμ-2–22 BCL-2 stimulated with goat F(ab′)2 anti-mouse IgM (10 μg/ml) for different times. β-Actin was used as control. The blot is representative of three independent experiments. (b) Analysis of Rasa3 expression and Erk1/2 and BimEL phosphorylation after splenic B cells activation with goat F(ab′)2 anti-mouse IgM (10 μg/ml) for different times or ionomycin (50 ng/ml) plus phorbol 12-myristate 13-acetate (10 ng/ml) for 5 min (I+P). Erk1/2 and β-actin were used as controls. The blot represents one of three independent experiments. (c) Rasa3–GFP subcellular distribution in Cos-7 cells transfected with Rasa3-GFP alone (Upper) or with Rasa3-GFP and Itpkb (Lower). Pictures of representative cells are presented 0, 3, and 5 min after the addition of 100 μM ATP. The profile of fluorescence intensity from membrane to membrane (as depicted by axis presented in small pictures at t = 0) is given below each cell. The fluorescence intensity is represented as arbitrary units. The ratio (R) between the mean fluorescence intensities of the membranes (A1 and A2) and the mean fluorescence intensity of cytosol (B) is given. The pictures are representative of three independent transfection experiments. (d) Ins(1,3,4,5)P4 decreases membrane localization of Rasa3–GFP in HeLa cells. Rasa3–GFP-transfected HeLa cells were cultured for 30 min with DMSO-containing solvent, <t>2,6-di-O-butyryl-Ins(1,2,4,5)P4/PM</t> (50 μg/ml) or 2,6-di-O-butyryl-Ins(1,3,4,5)P4/AM (50 μg/ml). Pictures of representative cells are presented 0 and 30 min after addition of each compound. Data are representative of three independent experiments.
Bio Gel P 4 Columns, supplied by Bio-Rad, 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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bio gel p4 polyacrylamide columns  (Bio-Rad)
97
Bio-Rad bio gel p4 polyacrylamide columns
Analysis of the signaling pathway leading to Bim increased expression. (a) Western blot for Bim expression in splenic B cells from Itpkb+/+ Eμ-2–22 BCL-2 and Itpkb−/− Eμ-2–22 BCL-2 stimulated with goat F(ab′)2 anti-mouse IgM (10 μg/ml) for different times. β-Actin was used as control. The blot is representative of three independent experiments. (b) Analysis of Rasa3 expression and Erk1/2 and BimEL phosphorylation after splenic B cells activation with goat F(ab′)2 anti-mouse IgM (10 μg/ml) for different times or ionomycin (50 ng/ml) plus phorbol 12-myristate 13-acetate (10 ng/ml) for 5 min (I+P). Erk1/2 and β-actin were used as controls. The blot represents one of three independent experiments. (c) Rasa3–GFP subcellular distribution in Cos-7 cells transfected with Rasa3-GFP alone (Upper) or with Rasa3-GFP and Itpkb (Lower). Pictures of representative cells are presented 0, 3, and 5 min after the addition of 100 μM ATP. The profile of fluorescence intensity from membrane to membrane (as depicted by axis presented in small pictures at t = 0) is given below each cell. The fluorescence intensity is represented as arbitrary units. The ratio (R) between the mean fluorescence intensities of the membranes (A1 and A2) and the mean fluorescence intensity of cytosol (B) is given. The pictures are representative of three independent transfection experiments. (d) Ins(1,3,4,5)P4 decreases membrane localization of Rasa3–GFP in HeLa cells. Rasa3–GFP-transfected HeLa cells were cultured for 30 min with DMSO-containing solvent, <t>2,6-di-O-butyryl-Ins(1,2,4,5)P4/PM</t> (50 μg/ml) or 2,6-di-O-butyryl-Ins(1,3,4,5)P4/AM (50 μg/ml). Pictures of representative cells are presented 0 and 30 min after addition of each compound. Data are representative of three independent experiments.
Bio Gel P4 Polyacrylamide Columns, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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bio gel p 4 micro spin buffer exchange columns  (Bio-Rad)
96
Bio-Rad bio gel p 4 micro spin buffer exchange columns
Analysis of the signaling pathway leading to Bim increased expression. (a) Western blot for Bim expression in splenic B cells from Itpkb+/+ Eμ-2–22 BCL-2 and Itpkb−/− Eμ-2–22 BCL-2 stimulated with goat F(ab′)2 anti-mouse IgM (10 μg/ml) for different times. β-Actin was used as control. The blot is representative of three independent experiments. (b) Analysis of Rasa3 expression and Erk1/2 and BimEL phosphorylation after splenic B cells activation with goat F(ab′)2 anti-mouse IgM (10 μg/ml) for different times or ionomycin (50 ng/ml) plus phorbol 12-myristate 13-acetate (10 ng/ml) for 5 min (I+P). Erk1/2 and β-actin were used as controls. The blot represents one of three independent experiments. (c) Rasa3–GFP subcellular distribution in Cos-7 cells transfected with Rasa3-GFP alone (Upper) or with Rasa3-GFP and Itpkb (Lower). Pictures of representative cells are presented 0, 3, and 5 min after the addition of 100 μM ATP. The profile of fluorescence intensity from membrane to membrane (as depicted by axis presented in small pictures at t = 0) is given below each cell. The fluorescence intensity is represented as arbitrary units. The ratio (R) between the mean fluorescence intensities of the membranes (A1 and A2) and the mean fluorescence intensity of cytosol (B) is given. The pictures are representative of three independent transfection experiments. (d) Ins(1,3,4,5)P4 decreases membrane localization of Rasa3–GFP in HeLa cells. Rasa3–GFP-transfected HeLa cells were cultured for 30 min with DMSO-containing solvent, <t>2,6-di-O-butyryl-Ins(1,2,4,5)P4/PM</t> (50 μg/ml) or 2,6-di-O-butyryl-Ins(1,3,4,5)P4/AM (50 μg/ml). Pictures of representative cells are presented 0 and 30 min after addition of each compound. Data are representative of three independent experiments.
Bio Gel P 4 Micro Spin Buffer Exchange Columns, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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chromatography column  (Bio-Rad)
93
Bio-Rad chromatography column
Analysis of the signaling pathway leading to Bim increased expression. (a) Western blot for Bim expression in splenic B cells from Itpkb+/+ Eμ-2–22 BCL-2 and Itpkb−/− Eμ-2–22 BCL-2 stimulated with goat F(ab′)2 anti-mouse IgM (10 μg/ml) for different times. β-Actin was used as control. The blot is representative of three independent experiments. (b) Analysis of Rasa3 expression and Erk1/2 and BimEL phosphorylation after splenic B cells activation with goat F(ab′)2 anti-mouse IgM (10 μg/ml) for different times or ionomycin (50 ng/ml) plus phorbol 12-myristate 13-acetate (10 ng/ml) for 5 min (I+P). Erk1/2 and β-actin were used as controls. The blot represents one of three independent experiments. (c) Rasa3–GFP subcellular distribution in Cos-7 cells transfected with Rasa3-GFP alone (Upper) or with Rasa3-GFP and Itpkb (Lower). Pictures of representative cells are presented 0, 3, and 5 min after the addition of 100 μM ATP. The profile of fluorescence intensity from membrane to membrane (as depicted by axis presented in small pictures at t = 0) is given below each cell. The fluorescence intensity is represented as arbitrary units. The ratio (R) between the mean fluorescence intensities of the membranes (A1 and A2) and the mean fluorescence intensity of cytosol (B) is given. The pictures are representative of three independent transfection experiments. (d) Ins(1,3,4,5)P4 decreases membrane localization of Rasa3–GFP in HeLa cells. Rasa3–GFP-transfected HeLa cells were cultured for 30 min with DMSO-containing solvent, <t>2,6-di-O-butyryl-Ins(1,2,4,5)P4/PM</t> (50 μg/ml) or 2,6-di-O-butyryl-Ins(1,3,4,5)P4/AM (50 μg/ml). Pictures of representative cells are presented 0 and 30 min after addition of each compound. Data are representative of three independent experiments.
Chromatography Column, supplied by Bio-Rad, 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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ins  (Echelon Biosciences)
93
Echelon Biosciences ins
Analysis of the signaling pathway leading to Bim increased expression. (a) Western blot for Bim expression in splenic B cells from Itpkb+/+ Eμ-2–22 BCL-2 and Itpkb−/− Eμ-2–22 BCL-2 stimulated with goat F(ab′)2 anti-mouse IgM (10 μg/ml) for different times. β-Actin was used as control. The blot is representative of three independent experiments. (b) Analysis of Rasa3 expression and Erk1/2 and BimEL phosphorylation after splenic B cells activation with goat F(ab′)2 anti-mouse IgM (10 μg/ml) for different times or ionomycin (50 ng/ml) plus phorbol 12-myristate 13-acetate (10 ng/ml) for 5 min (I+P). Erk1/2 and β-actin were used as controls. The blot represents one of three independent experiments. (c) Rasa3–GFP subcellular distribution in Cos-7 cells transfected with Rasa3-GFP alone (Upper) or with Rasa3-GFP and Itpkb (Lower). Pictures of representative cells are presented 0, 3, and 5 min after the addition of 100 μM ATP. The profile of fluorescence intensity from membrane to membrane (as depicted by axis presented in small pictures at t = 0) is given below each cell. The fluorescence intensity is represented as arbitrary units. The ratio (R) between the mean fluorescence intensities of the membranes (A1 and A2) and the mean fluorescence intensity of cytosol (B) is given. The pictures are representative of three independent transfection experiments. (d) Ins(1,3,4,5)P4 decreases membrane localization of Rasa3–GFP in HeLa cells. Rasa3–GFP-transfected HeLa cells were cultured for 30 min with DMSO-containing solvent, <t>2,6-di-O-butyryl-Ins(1,2,4,5)P4/PM</t> (50 μg/ml) or 2,6-di-O-butyryl-Ins(1,3,4,5)P4/AM (50 μg/ml). Pictures of representative cells are presented 0 and 30 min after addition of each compound. Data are representative of three independent experiments.
Ins, supplied by Echelon Biosciences, 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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d - threo -p4 (p4  (Matreya LLC)
90
Matreya LLC d - threo -p4 (p4
Analysis of the signaling pathway leading to Bim increased expression. (a) Western blot for Bim expression in splenic B cells from Itpkb+/+ Eμ-2–22 BCL-2 and Itpkb−/− Eμ-2–22 BCL-2 stimulated with goat F(ab′)2 anti-mouse IgM (10 μg/ml) for different times. β-Actin was used as control. The blot is representative of three independent experiments. (b) Analysis of Rasa3 expression and Erk1/2 and BimEL phosphorylation after splenic B cells activation with goat F(ab′)2 anti-mouse IgM (10 μg/ml) for different times or ionomycin (50 ng/ml) plus phorbol 12-myristate 13-acetate (10 ng/ml) for 5 min (I+P). Erk1/2 and β-actin were used as controls. The blot represents one of three independent experiments. (c) Rasa3–GFP subcellular distribution in Cos-7 cells transfected with Rasa3-GFP alone (Upper) or with Rasa3-GFP and Itpkb (Lower). Pictures of representative cells are presented 0, 3, and 5 min after the addition of 100 μM ATP. The profile of fluorescence intensity from membrane to membrane (as depicted by axis presented in small pictures at t = 0) is given below each cell. The fluorescence intensity is represented as arbitrary units. The ratio (R) between the mean fluorescence intensities of the membranes (A1 and A2) and the mean fluorescence intensity of cytosol (B) is given. The pictures are representative of three independent transfection experiments. (d) Ins(1,3,4,5)P4 decreases membrane localization of Rasa3–GFP in HeLa cells. Rasa3–GFP-transfected HeLa cells were cultured for 30 min with DMSO-containing solvent, <t>2,6-di-O-butyryl-Ins(1,2,4,5)P4/PM</t> (50 μg/ml) or 2,6-di-O-butyryl-Ins(1,3,4,5)P4/AM (50 μg/ml). Pictures of representative cells are presented 0 and 30 min after addition of each compound. Data are representative of three independent experiments.
D Threo P4 (P4, supplied by Matreya LLC, 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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Analysis of the signaling pathway leading to Bim increased expression. (a) Western blot for Bim expression in splenic B cells from Itpkb+/+ Eμ-2–22 BCL-2 and Itpkb−/− Eμ-2–22 BCL-2 stimulated with goat F(ab′)2 anti-mouse IgM (10 μg/ml) for different times. β-Actin was used as control. The blot is representative of three independent experiments. (b) Analysis of Rasa3 expression and Erk1/2 and BimEL phosphorylation after splenic B cells activation with goat F(ab′)2 anti-mouse IgM (10 μg/ml) for different times or ionomycin (50 ng/ml) plus phorbol 12-myristate 13-acetate (10 ng/ml) for 5 min (I+P). Erk1/2 and β-actin were used as controls. The blot represents one of three independent experiments. (c) Rasa3–GFP subcellular distribution in Cos-7 cells transfected with Rasa3-GFP alone (Upper) or with Rasa3-GFP and Itpkb (Lower). Pictures of representative cells are presented 0, 3, and 5 min after the addition of 100 μM ATP. The profile of fluorescence intensity from membrane to membrane (as depicted by axis presented in small pictures at t = 0) is given below each cell. The fluorescence intensity is represented as arbitrary units. The ratio (R) between the mean fluorescence intensities of the membranes (A1 and A2) and the mean fluorescence intensity of cytosol (B) is given. The pictures are representative of three independent transfection experiments. (d) Ins(1,3,4,5)P4 decreases membrane localization of Rasa3–GFP in HeLa cells. Rasa3–GFP-transfected HeLa cells were cultured for 30 min with DMSO-containing solvent, 2,6-di-O-butyryl-Ins(1,2,4,5)P4/PM (50 μg/ml) or 2,6-di-O-butyryl-Ins(1,3,4,5)P4/AM (50 μg/ml). Pictures of representative cells are presented 0 and 30 min after addition of each compound. Data are representative of three independent experiments.

Journal:

Article Title: Inositol 1,3,4,5-tetrakisphosphate controls proapoptotic Bim gene expression and survival in B cells

doi: 10.1073/pnas.0704312104

Figure Lengend Snippet: Analysis of the signaling pathway leading to Bim increased expression. (a) Western blot for Bim expression in splenic B cells from Itpkb+/+ Eμ-2–22 BCL-2 and Itpkb−/− Eμ-2–22 BCL-2 stimulated with goat F(ab′)2 anti-mouse IgM (10 μg/ml) for different times. β-Actin was used as control. The blot is representative of three independent experiments. (b) Analysis of Rasa3 expression and Erk1/2 and BimEL phosphorylation after splenic B cells activation with goat F(ab′)2 anti-mouse IgM (10 μg/ml) for different times or ionomycin (50 ng/ml) plus phorbol 12-myristate 13-acetate (10 ng/ml) for 5 min (I+P). Erk1/2 and β-actin were used as controls. The blot represents one of three independent experiments. (c) Rasa3–GFP subcellular distribution in Cos-7 cells transfected with Rasa3-GFP alone (Upper) or with Rasa3-GFP and Itpkb (Lower). Pictures of representative cells are presented 0, 3, and 5 min after the addition of 100 μM ATP. The profile of fluorescence intensity from membrane to membrane (as depicted by axis presented in small pictures at t = 0) is given below each cell. The fluorescence intensity is represented as arbitrary units. The ratio (R) between the mean fluorescence intensities of the membranes (A1 and A2) and the mean fluorescence intensity of cytosol (B) is given. The pictures are representative of three independent transfection experiments. (d) Ins(1,3,4,5)P4 decreases membrane localization of Rasa3–GFP in HeLa cells. Rasa3–GFP-transfected HeLa cells were cultured for 30 min with DMSO-containing solvent, 2,6-di-O-butyryl-Ins(1,2,4,5)P4/PM (50 μg/ml) or 2,6-di-O-butyryl-Ins(1,3,4,5)P4/AM (50 μg/ml). Pictures of representative cells are presented 0 and 30 min after addition of each compound. Data are representative of three independent experiments.

Article Snippet: The isoform Ins(1,2,4,5) P 4 , Bt 2 Ins(1,2,4,5) P 4 /PM (Echelon Bioscience) and DMSO solvant were used as controls.

Techniques: Expressing, Western Blot, Activation Assay, Transfection, Fluorescence, Cell Culture