phospho-mek1 Search Results


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  • 98
    Millipore anti phospho mek1
    Anti Phospho Mek1, supplied by Millipore, used in various techniques. Bioz Stars score: 98/100, based on 23 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Cell Signaling Technology Inc phospho mek1 2
    Phospho Mek1 2, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 99/100, based on 809 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Cell Signaling Technology Inc anti phospho mek1 2
    Anti Phospho Mek1 2, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 99/100, based on 286 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    97
    Cell Signaling Technology Inc phosphorylated mek1 2
    Elevated miR-30d suppresses the MEK/ERK and PI3K/Akt signaling pathways in SK-ES-1 cells. (A) Western blot assay revealed that overexpression of miR-30d reduced the expression levels of <t>p-MEK1/2</t> and p-ERK1/2, but caused no changes in the levels of MEK1/2 and ERK1/2. (B) Ratios of p-MEK/MEK and p-ERK/ERK in miR-30d mimic group were decreased compared with those in the untreated group (**P
    Phosphorylated Mek1 2, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 97/100, based on 186 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Cell Signaling Technology Inc mek1 2
    Elevated miR-30d suppresses the MEK/ERK and PI3K/Akt signaling pathways in SK-ES-1 cells. (A) Western blot assay revealed that overexpression of miR-30d reduced the expression levels of <t>p-MEK1/2</t> and p-ERK1/2, but caused no changes in the levels of MEK1/2 and ERK1/2. (B) Ratios of p-MEK/MEK and p-ERK/ERK in miR-30d mimic group were decreased compared with those in the untreated group (**P
    Mek1 2, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 99/100, based on 2296 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Cell Signaling Technology Inc rabbit anti phospho mek1 2
    Elevated miR-30d suppresses the MEK/ERK and PI3K/Akt signaling pathways in SK-ES-1 cells. (A) Western blot assay revealed that overexpression of miR-30d reduced the expression levels of <t>p-MEK1/2</t> and p-ERK1/2, but caused no changes in the levels of MEK1/2 and ERK1/2. (B) Ratios of p-MEK/MEK and p-ERK/ERK in miR-30d mimic group were decreased compared with those in the untreated group (**P
    Rabbit Anti Phospho Mek1 2, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 88/100, based on 84 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    88
    Cell Signaling Technology Inc phospho mek1
    MEKK2 deficiency partially reverses NF1-associated skeletal pathology in mice. a Femurs from 16-week-old Nf1 fl/fl , Nf1 fl/fl ; Mekk2 −/− , Nf1 fl/fl ; Dmp1-Cre , and Nf1 fl/fl ; Mekk2 −/− ; Dmp1-Cre mice were analyzed by μCT. b Quantitative parameters are porous volume, porosity (porous volume/total volume), and porous surface from 16-week-old Nf1 fl/fl ( n = 5), Nf1 fl/fl ; Mekk2 −/− ( n = 8), Nf1 fl/fl ; Dmp1-Cre ( n = 8), and Nf1 fl/fl ; Mekk2 −/− ; Dmp1-Cre ( n = 10) mice. Mean ± s.d., one-way ANOVA with Tukey’s multiple comparison test. c Representative μCT 3D-reconstuction images of trabecular bone in the distal femur metaphysis. d Quantitative parameters include trabecular bone volume/total volume (BV/TV), thickness (Tb.Th), trabecular number (Tb.N), and spacing (Tb.sp) in 16-week-old Nf1 fl/fl ( n = 8), Nf1 fl/fl ; Mekk2 −/− ( n = 6), Nf1 fl/fl ; Dmp1-Cre ( n = 9), and Nf1 fl/fl ; Mekk2 −/− ; Dmp1-Cre ( n = 8) mice. mean ± s.d., one-way ANOVA with Tukey’s multiple comparison test. e μCT scans of mouse skulls at 16 weeks Nf1 fl/fl , Nf1 fl/fl ; Mekk2 −/− , Nf1 fl/fl ; Dmp1-Cre , and Nf1 fl/fl ; Mekk2 −/− ; Dmp1-Cre mice. f Serum levels of P1NP, CTX, FGF23, PTH, 25(OH) vitamin D, and phosphate in 16-week-old mice. Data are represented as box plots with the middle line representing the median, the box representing the 95% confidence interval of the median, and the whiskers representing the range. Each dot represents a separate mouse. g Representative images and quantification of immunostaining for p-MEKK2 (green), <t>p-MEK1</t> (magenta), and p-ERK (red) in trabecular (left panels) and cortical bone (right panels) from 16-week-old Nf1 fl/fl , Nf1 fl/fl ; Dmp1-Cre , and Nf1 fl/fl ; Mekk2 −/− ; Dmp1-Cre mice. Nuclei are counterstained with DAPI (blue) and the scale bar indicates 100 µm. Three independent fields were examined per mouse ( n = 3 mice per group). mean ± s.d., one-way ANOVA with Tukey’s multiple comparison test. * P
    Phospho Mek1, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 88/100, based on 45 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    97
    Abcam anti mek1 phospho s298 antibody epr3338
    TBMS1 inactivates VEGFR2 mediated ERK pathway. (A) RT-PCR analysis of VEGFR-2, <t>MEK1</t> and ERK mRNA expression levels. β-actin was used as an internal control. (B) Western blot analysis on VEGFR-2 protein expression and MEK1 and ERK1/2 phosphorylation levels. GADPH was served as an internal control. Representative blots are presented with corresponding densitometric analysis is shown. Data are given as mean ± SD from three independent experiments. Compared with control, **P
    Anti Mek1 Phospho S298 Antibody Epr3338, supplied by Abcam, used in various techniques. Bioz Stars score: 97/100, based on 21 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Cell Signaling Technology Inc rabbit anti phospho mek1 2 ser217 221
    TBMS1 inactivates VEGFR2 mediated ERK pathway. (A) RT-PCR analysis of VEGFR-2, <t>MEK1</t> and ERK mRNA expression levels. β-actin was used as an internal control. (B) Western blot analysis on VEGFR-2 protein expression and MEK1 and ERK1/2 phosphorylation levels. GADPH was served as an internal control. Representative blots are presented with corresponding densitometric analysis is shown. Data are given as mean ± SD from three independent experiments. Compared with control, **P
    Rabbit Anti Phospho Mek1 2 Ser217 221, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 94/100, based on 37 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Cell Signaling Technology Inc phospho mek1 2 s217 221
    TBMS1 inactivates VEGFR2 mediated ERK pathway. (A) RT-PCR analysis of VEGFR-2, <t>MEK1</t> and ERK mRNA expression levels. β-actin was used as an internal control. (B) Western blot analysis on VEGFR-2 protein expression and MEK1 and ERK1/2 phosphorylation levels. GADPH was served as an internal control. Representative blots are presented with corresponding densitometric analysis is shown. Data are given as mean ± SD from three independent experiments. Compared with control, **P
    Phospho Mek1 2 S217 221, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 97/100, based on 36 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Cell Signaling Technology Inc anti phospho mek1
    <t>MEK1</t> is required to inhibit the expression of the pluripotency genes pou5f3.2 and ventx2 . ( A ) Embryos injected with 25 ng Mk-MO at 16-cell stage in one animal dorsal blastomere were grown until late gastrulation stage 13 and processed for WISH with pou5f3.2 and ventx2 probes. ( B ) Embryos injected with 25 ng Mk-MO at 16 cell stage in one animal ventral blastomere were grown until mid-neurula stage 18 and processed for WISH with pou5f3.2 probe. ( C ) Four-cell embryos were injected in each blastomere with 25 ng Mk-MO and grown until blastula stage 9, when animal caps were isolated, cultured in vitro until late gastrula stage 13 and then processed for RT-qPCR. In A and B, the number of embryos exemplified by the photograph over the total number of embryos analyzed is indicated. DOI: http://dx.doi.org/10.7554/eLife.21526.008
    Anti Phospho Mek1, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 99/100, based on 13 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Cell Signaling Technology Inc phospho mek1 2 ser221
    <t>MEK1</t> is required to inhibit the expression of the pluripotency genes pou5f3.2 and ventx2 . ( A ) Embryos injected with 25 ng Mk-MO at 16-cell stage in one animal dorsal blastomere were grown until late gastrulation stage 13 and processed for WISH with pou5f3.2 and ventx2 probes. ( B ) Embryos injected with 25 ng Mk-MO at 16 cell stage in one animal ventral blastomere were grown until mid-neurula stage 18 and processed for WISH with pou5f3.2 probe. ( C ) Four-cell embryos were injected in each blastomere with 25 ng Mk-MO and grown until blastula stage 9, when animal caps were isolated, cultured in vitro until late gastrula stage 13 and then processed for RT-qPCR. In A and B, the number of embryos exemplified by the photograph over the total number of embryos analyzed is indicated. DOI: http://dx.doi.org/10.7554/eLife.21526.008
    Phospho Mek1 2 Ser221, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 99/100, based on 22 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Cell Signaling Technology Inc phospho mek1 2 ser217 221 rabbit
    <t>MEK1</t> is required to inhibit the expression of the pluripotency genes pou5f3.2 and ventx2 . ( A ) Embryos injected with 25 ng Mk-MO at 16-cell stage in one animal dorsal blastomere were grown until late gastrulation stage 13 and processed for WISH with pou5f3.2 and ventx2 probes. ( B ) Embryos injected with 25 ng Mk-MO at 16 cell stage in one animal ventral blastomere were grown until mid-neurula stage 18 and processed for WISH with pou5f3.2 probe. ( C ) Four-cell embryos were injected in each blastomere with 25 ng Mk-MO and grown until blastula stage 9, when animal caps were isolated, cultured in vitro until late gastrula stage 13 and then processed for RT-qPCR. In A and B, the number of embryos exemplified by the photograph over the total number of embryos analyzed is indicated. DOI: http://dx.doi.org/10.7554/eLife.21526.008
    Phospho Mek1 2 Ser217 221 Rabbit, supplied by Cell Signaling Technology Inc, 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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    Abcam anti p mek1 2
    <t>MEK1</t> is required to inhibit the expression of the pluripotency genes pou5f3.2 and ventx2 . ( A ) Embryos injected with 25 ng Mk-MO at 16-cell stage in one animal dorsal blastomere were grown until late gastrulation stage 13 and processed for WISH with pou5f3.2 and ventx2 probes. ( B ) Embryos injected with 25 ng Mk-MO at 16 cell stage in one animal ventral blastomere were grown until mid-neurula stage 18 and processed for WISH with pou5f3.2 probe. ( C ) Four-cell embryos were injected in each blastomere with 25 ng Mk-MO and grown until blastula stage 9, when animal caps were isolated, cultured in vitro until late gastrula stage 13 and then processed for RT-qPCR. In A and B, the number of embryos exemplified by the photograph over the total number of embryos analyzed is indicated. DOI: http://dx.doi.org/10.7554/eLife.21526.008
    Anti P Mek1 2, supplied by Abcam, used in various techniques. Bioz Stars score: 94/100, based on 21 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Elevated miR-30d suppresses the MEK/ERK and PI3K/Akt signaling pathways in SK-ES-1 cells. (A) Western blot assay revealed that overexpression of miR-30d reduced the expression levels of p-MEK1/2 and p-ERK1/2, but caused no changes in the levels of MEK1/2 and ERK1/2. (B) Ratios of p-MEK/MEK and p-ERK/ERK in miR-30d mimic group were decreased compared with those in the untreated group (**P

    Journal: Oncology Letters

    Article Title: miR-30d inhibits cell biological progression of Ewing's sarcoma by suppressing the MEK/ERK and PI3K/Akt pathways in vitro

    doi: 10.3892/ol.2018.7900

    Figure Lengend Snippet: Elevated miR-30d suppresses the MEK/ERK and PI3K/Akt signaling pathways in SK-ES-1 cells. (A) Western blot assay revealed that overexpression of miR-30d reduced the expression levels of p-MEK1/2 and p-ERK1/2, but caused no changes in the levels of MEK1/2 and ERK1/2. (B) Ratios of p-MEK/MEK and p-ERK/ERK in miR-30d mimic group were decreased compared with those in the untreated group (**P

    Article Snippet: Antibodies against MMP-2 (cat. no. 40994), MMP-9 (cat. no. 13667), Bax (cat. no. 5023), Bcl-2 (cat. no. 4223), caspase-3 (cat. no. 9665), PARP (cat. no. 9532), MEK1/2 (cat. no. 8727), ERK1/2 (cat. no. 4695), phosphorylated MEK1/2 (p-MEK1/2) (cat. no. 9154), phosphorylated ERK1/2 (p-ERK1/2) (cat. no. 4370), phosphoinositide 3-kinase (PI3K) (cat. no. 4249), Akt (cat. no. 4691), phosphorylated Akt (p-Akt) (cat. no. 4060) and β-actin (cat. no. 4970) were purchased from Cell Signaling Technology, Inc. (Danvers, MA, USA).

    Techniques: Western Blot, Over Expression, Expressing

    MEKK2 deficiency partially reverses NF1-associated skeletal pathology in mice. a Femurs from 16-week-old Nf1 fl/fl , Nf1 fl/fl ; Mekk2 −/− , Nf1 fl/fl ; Dmp1-Cre , and Nf1 fl/fl ; Mekk2 −/− ; Dmp1-Cre mice were analyzed by μCT. b Quantitative parameters are porous volume, porosity (porous volume/total volume), and porous surface from 16-week-old Nf1 fl/fl ( n = 5), Nf1 fl/fl ; Mekk2 −/− ( n = 8), Nf1 fl/fl ; Dmp1-Cre ( n = 8), and Nf1 fl/fl ; Mekk2 −/− ; Dmp1-Cre ( n = 10) mice. Mean ± s.d., one-way ANOVA with Tukey’s multiple comparison test. c Representative μCT 3D-reconstuction images of trabecular bone in the distal femur metaphysis. d Quantitative parameters include trabecular bone volume/total volume (BV/TV), thickness (Tb.Th), trabecular number (Tb.N), and spacing (Tb.sp) in 16-week-old Nf1 fl/fl ( n = 8), Nf1 fl/fl ; Mekk2 −/− ( n = 6), Nf1 fl/fl ; Dmp1-Cre ( n = 9), and Nf1 fl/fl ; Mekk2 −/− ; Dmp1-Cre ( n = 8) mice. mean ± s.d., one-way ANOVA with Tukey’s multiple comparison test. e μCT scans of mouse skulls at 16 weeks Nf1 fl/fl , Nf1 fl/fl ; Mekk2 −/− , Nf1 fl/fl ; Dmp1-Cre , and Nf1 fl/fl ; Mekk2 −/− ; Dmp1-Cre mice. f Serum levels of P1NP, CTX, FGF23, PTH, 25(OH) vitamin D, and phosphate in 16-week-old mice. Data are represented as box plots with the middle line representing the median, the box representing the 95% confidence interval of the median, and the whiskers representing the range. Each dot represents a separate mouse. g Representative images and quantification of immunostaining for p-MEKK2 (green), p-MEK1 (magenta), and p-ERK (red) in trabecular (left panels) and cortical bone (right panels) from 16-week-old Nf1 fl/fl , Nf1 fl/fl ; Dmp1-Cre , and Nf1 fl/fl ; Mekk2 −/− ; Dmp1-Cre mice. Nuclei are counterstained with DAPI (blue) and the scale bar indicates 100 µm. Three independent fields were examined per mouse ( n = 3 mice per group). mean ± s.d., one-way ANOVA with Tukey’s multiple comparison test. * P

    Journal: Nature Communications

    Article Title: MEKK2 mediates aberrant ERK activation in neurofibromatosis type I

    doi: 10.1038/s41467-020-19555-6

    Figure Lengend Snippet: MEKK2 deficiency partially reverses NF1-associated skeletal pathology in mice. a Femurs from 16-week-old Nf1 fl/fl , Nf1 fl/fl ; Mekk2 −/− , Nf1 fl/fl ; Dmp1-Cre , and Nf1 fl/fl ; Mekk2 −/− ; Dmp1-Cre mice were analyzed by μCT. b Quantitative parameters are porous volume, porosity (porous volume/total volume), and porous surface from 16-week-old Nf1 fl/fl ( n = 5), Nf1 fl/fl ; Mekk2 −/− ( n = 8), Nf1 fl/fl ; Dmp1-Cre ( n = 8), and Nf1 fl/fl ; Mekk2 −/− ; Dmp1-Cre ( n = 10) mice. Mean ± s.d., one-way ANOVA with Tukey’s multiple comparison test. c Representative μCT 3D-reconstuction images of trabecular bone in the distal femur metaphysis. d Quantitative parameters include trabecular bone volume/total volume (BV/TV), thickness (Tb.Th), trabecular number (Tb.N), and spacing (Tb.sp) in 16-week-old Nf1 fl/fl ( n = 8), Nf1 fl/fl ; Mekk2 −/− ( n = 6), Nf1 fl/fl ; Dmp1-Cre ( n = 9), and Nf1 fl/fl ; Mekk2 −/− ; Dmp1-Cre ( n = 8) mice. mean ± s.d., one-way ANOVA with Tukey’s multiple comparison test. e μCT scans of mouse skulls at 16 weeks Nf1 fl/fl , Nf1 fl/fl ; Mekk2 −/− , Nf1 fl/fl ; Dmp1-Cre , and Nf1 fl/fl ; Mekk2 −/− ; Dmp1-Cre mice. f Serum levels of P1NP, CTX, FGF23, PTH, 25(OH) vitamin D, and phosphate in 16-week-old mice. Data are represented as box plots with the middle line representing the median, the box representing the 95% confidence interval of the median, and the whiskers representing the range. Each dot represents a separate mouse. g Representative images and quantification of immunostaining for p-MEKK2 (green), p-MEK1 (magenta), and p-ERK (red) in trabecular (left panels) and cortical bone (right panels) from 16-week-old Nf1 fl/fl , Nf1 fl/fl ; Dmp1-Cre , and Nf1 fl/fl ; Mekk2 −/− ; Dmp1-Cre mice. Nuclei are counterstained with DAPI (blue) and the scale bar indicates 100 µm. Three independent fields were examined per mouse ( n = 3 mice per group). mean ± s.d., one-way ANOVA with Tukey’s multiple comparison test. * P

    Article Snippet: Samples were then incubated with primary antibody against phospho-MEKK2 (Ser520; cat. no. PA5-105898, Invitrogen), phospho-MEK1 (Thr286; cat. no. 9127, Cell Signaling Technology), phospho-ERK1/2 (Thr202/Tyr204; cat. no. 4370, Cell Signaling Technology), phospho-AKT (Ser473; cat. no. 4060, Cell Signaling Technology) overnight at 4 °C, then washed three times with PBS.

    Techniques: Mouse Assay, Immunostaining

    Inhibition of MEKK2 ameliorates skeletal defects in a mouse model of skeletal NF1. a Femurs from 16-week-old Nf1 fl/fl mice treated with vehicle ( n = 4) or ponatinib ( n = 5) and Nf1 fl/fl ; Dmp1-Cre mice treated with vehicle ( n = 7) or ponatinib ( n = 6) were analyzed by μCT. b Quantitative parameters are porous volume, porosity (porous volume/total volume), and porous surface from vehicle (veh) and ponatinib-treated groups (pon) of Nf1 fl/fl ;Dmp1-Cre mice. Mean ± s.d., one-way ANOVA with Tukey’s multiple comparison test. c Representative μCT 3D-reconstruction images of trabecular bone in the distal femur metaphysis. d Relative quantitative analysis of trabecular BV/TV, thickness (Tb.Th), trabecular number (Tb.N), and spacing (Tb.sp) in vehicle ( n = 8) and ponatinib-treated ( n = 7) group of Nf1 fl/fl ;Dmp1-Cre . Mean ± s.d., unpaired, two-tailed Student’s t test. e μCT scans of mouse skulls in 16-week-old Nf1 fl/fl and Nf1 fl/fl ;Dmp1-Cre mice treated with vehicle or ponatinib. f Serum levels of PINP, 25(OH) vitamin D, and phosphate in 16-week-old Nf1 fl/fl and Nf1 fl/fl ;Dmp1-Cre mice treated with vehicle (veh) or ponatinib (pon). Data are represented as box plots with the middle line representing the median, the box representing the 95% confidence interval of the median, and the whiskers representing the range. Each dot represents a separate mouse. g Representative immunofluorescent images and quantification for p-MEKK2 (green), p-MEK1 (magenta), and p-ERK (red) in femurs from 16-week-old Nf1 fl/fl (WT) and Nf1 fl/fl ;Dmp1-Cre mice treated with vehicle or ponatinib. Far right images show enlarged views of the dotted orange boxes. White arrows indicate signal positive osteoblasts. Nuclei are counterstained with DAPI (blue) and the scale bar indicates 100 µm. Three independent fields were examined per mouse ( n = 3 mice per group). Mean ± s.d., one-way ANOVA with Tukey’s multiple comparison test. * P

    Journal: Nature Communications

    Article Title: MEKK2 mediates aberrant ERK activation in neurofibromatosis type I

    doi: 10.1038/s41467-020-19555-6

    Figure Lengend Snippet: Inhibition of MEKK2 ameliorates skeletal defects in a mouse model of skeletal NF1. a Femurs from 16-week-old Nf1 fl/fl mice treated with vehicle ( n = 4) or ponatinib ( n = 5) and Nf1 fl/fl ; Dmp1-Cre mice treated with vehicle ( n = 7) or ponatinib ( n = 6) were analyzed by μCT. b Quantitative parameters are porous volume, porosity (porous volume/total volume), and porous surface from vehicle (veh) and ponatinib-treated groups (pon) of Nf1 fl/fl ;Dmp1-Cre mice. Mean ± s.d., one-way ANOVA with Tukey’s multiple comparison test. c Representative μCT 3D-reconstruction images of trabecular bone in the distal femur metaphysis. d Relative quantitative analysis of trabecular BV/TV, thickness (Tb.Th), trabecular number (Tb.N), and spacing (Tb.sp) in vehicle ( n = 8) and ponatinib-treated ( n = 7) group of Nf1 fl/fl ;Dmp1-Cre . Mean ± s.d., unpaired, two-tailed Student’s t test. e μCT scans of mouse skulls in 16-week-old Nf1 fl/fl and Nf1 fl/fl ;Dmp1-Cre mice treated with vehicle or ponatinib. f Serum levels of PINP, 25(OH) vitamin D, and phosphate in 16-week-old Nf1 fl/fl and Nf1 fl/fl ;Dmp1-Cre mice treated with vehicle (veh) or ponatinib (pon). Data are represented as box plots with the middle line representing the median, the box representing the 95% confidence interval of the median, and the whiskers representing the range. Each dot represents a separate mouse. g Representative immunofluorescent images and quantification for p-MEKK2 (green), p-MEK1 (magenta), and p-ERK (red) in femurs from 16-week-old Nf1 fl/fl (WT) and Nf1 fl/fl ;Dmp1-Cre mice treated with vehicle or ponatinib. Far right images show enlarged views of the dotted orange boxes. White arrows indicate signal positive osteoblasts. Nuclei are counterstained with DAPI (blue) and the scale bar indicates 100 µm. Three independent fields were examined per mouse ( n = 3 mice per group). Mean ± s.d., one-way ANOVA with Tukey’s multiple comparison test. * P

    Article Snippet: Samples were then incubated with primary antibody against phospho-MEKK2 (Ser520; cat. no. PA5-105898, Invitrogen), phospho-MEK1 (Thr286; cat. no. 9127, Cell Signaling Technology), phospho-ERK1/2 (Thr202/Tyr204; cat. no. 4370, Cell Signaling Technology), phospho-AKT (Ser473; cat. no. 4060, Cell Signaling Technology) overnight at 4 °C, then washed three times with PBS.

    Techniques: Inhibition, Mouse Assay, Two Tailed Test

    Pharmacologic inhibition of MEKK2 prohibits ERK activation. a Primary COBs were immortalized through infection with a retrovirus expressing SV40 large T-Antigen. COBs were pre-treated with the indicated inhibitors (1 µM, 30 min), then stimulated with or without FGF2 (25 ng/ml, 20 min) 30 min later. MEKK2 phosphorylation was analyzed by phos-tag electrophoresis. b MEKK2 phosphorylation was analyzed by either phos-tag electrophoresis or immunoblotting with anti-p-MEKK2 in primary Nf1 fl/fl osteoblasts infected with either vec or Cre lentivirus with the indicated inhibitors and FGF2 stimulation. c hMSCs were treated with the indicated inhibitors, then MEKK2 phosphorylation was examined by phos-tag electrophoresis. d WT immortalized COBs were lysed and immunoblotted for the indicated antibodies after inhibitor treatment. e Activation of ERK was detected in Saos-2 cells after incubation with the indicated inhibitors and stimulation with or without FGF2. f Purified unactivated GST-MEK1 was incubated with purified GST-MEKK2 and the indicated doses of ponatinib, and kinase activity of MEKK2 was analyzed by an in vitro kinase assay. g Expression levels of osteoblast genes in primary Nf1 fl/fl osteoblasts infected with either vec or Cre lentivirus. After infection, these cells are treated with the indicated inhibitors or a Mekk2 -targeting shRNA and cultured for 14 days ( n = 4 biologically independent samples). mean ± s.d. h Saos-2 cells were treated with DMSO, ponatinib (1 µM), BRITE-0600690 (BRITE-690, inactive compound), and BRITE-0600719 (BRITE-719, active compound) for 1 h with or without FGF2 stimulation. Phosphorylation levels of ERK1/2 were assessed by immunoblotting. All data shown are representative of either two or three total independent experiments. All unprocessed blots are provided in Supplementary Fig. 5 . Source data are provided as a Source Data file.

    Journal: Nature Communications

    Article Title: MEKK2 mediates aberrant ERK activation in neurofibromatosis type I

    doi: 10.1038/s41467-020-19555-6

    Figure Lengend Snippet: Pharmacologic inhibition of MEKK2 prohibits ERK activation. a Primary COBs were immortalized through infection with a retrovirus expressing SV40 large T-Antigen. COBs were pre-treated with the indicated inhibitors (1 µM, 30 min), then stimulated with or without FGF2 (25 ng/ml, 20 min) 30 min later. MEKK2 phosphorylation was analyzed by phos-tag electrophoresis. b MEKK2 phosphorylation was analyzed by either phos-tag electrophoresis or immunoblotting with anti-p-MEKK2 in primary Nf1 fl/fl osteoblasts infected with either vec or Cre lentivirus with the indicated inhibitors and FGF2 stimulation. c hMSCs were treated with the indicated inhibitors, then MEKK2 phosphorylation was examined by phos-tag electrophoresis. d WT immortalized COBs were lysed and immunoblotted for the indicated antibodies after inhibitor treatment. e Activation of ERK was detected in Saos-2 cells after incubation with the indicated inhibitors and stimulation with or without FGF2. f Purified unactivated GST-MEK1 was incubated with purified GST-MEKK2 and the indicated doses of ponatinib, and kinase activity of MEKK2 was analyzed by an in vitro kinase assay. g Expression levels of osteoblast genes in primary Nf1 fl/fl osteoblasts infected with either vec or Cre lentivirus. After infection, these cells are treated with the indicated inhibitors or a Mekk2 -targeting shRNA and cultured for 14 days ( n = 4 biologically independent samples). mean ± s.d. h Saos-2 cells were treated with DMSO, ponatinib (1 µM), BRITE-0600690 (BRITE-690, inactive compound), and BRITE-0600719 (BRITE-719, active compound) for 1 h with or without FGF2 stimulation. Phosphorylation levels of ERK1/2 were assessed by immunoblotting. All data shown are representative of either two or three total independent experiments. All unprocessed blots are provided in Supplementary Fig. 5 . Source data are provided as a Source Data file.

    Article Snippet: Samples were then incubated with primary antibody against phospho-MEKK2 (Ser520; cat. no. PA5-105898, Invitrogen), phospho-MEK1 (Thr286; cat. no. 9127, Cell Signaling Technology), phospho-ERK1/2 (Thr202/Tyr204; cat. no. 4370, Cell Signaling Technology), phospho-AKT (Ser473; cat. no. 4060, Cell Signaling Technology) overnight at 4 °C, then washed three times with PBS.

    Techniques: Inhibition, Activation Assay, Infection, Expressing, Electrophoresis, Incubation, Purification, Activity Assay, In Vitro, Kinase Assay, shRNA, Cell Culture

    TBMS1 inactivates VEGFR2 mediated ERK pathway. (A) RT-PCR analysis of VEGFR-2, MEK1 and ERK mRNA expression levels. β-actin was used as an internal control. (B) Western blot analysis on VEGFR-2 protein expression and MEK1 and ERK1/2 phosphorylation levels. GADPH was served as an internal control. Representative blots are presented with corresponding densitometric analysis is shown. Data are given as mean ± SD from three independent experiments. Compared with control, **P

    Journal: Oncology Letters

    Article Title: Tubeimoside-1 inhibits the proliferation and metastasis by promoting miR-126-5p expression in non-small cell lung cancer cellsNF-kB, JNK and p53 pathways are involved in tubeimoside-1-induced apoptosis in HepG2 cells with oxidative stress and G (

    doi: 10.3892/ol.2018.9051

    Figure Lengend Snippet: TBMS1 inactivates VEGFR2 mediated ERK pathway. (A) RT-PCR analysis of VEGFR-2, MEK1 and ERK mRNA expression levels. β-actin was used as an internal control. (B) Western blot analysis on VEGFR-2 protein expression and MEK1 and ERK1/2 phosphorylation levels. GADPH was served as an internal control. Representative blots are presented with corresponding densitometric analysis is shown. Data are given as mean ± SD from three independent experiments. Compared with control, **P

    Article Snippet: The blots were then blocked with 5% non-fat milk overnight at 4°C and incubated at 4°C overnight with the specific primary antibodies as follows: anti-VEGFR-2, anti-ERK1/2, anti-p-ERK1 (pT202/pY204)+p-ERK2 (pT185/pY187) (all 1:1,500 diluted; Abcam, Cambridge, MA, USA); anti-MEK1, anti-p-MEK1 (pS298) (both 1:1,000 diluted; Abcam).

    Techniques: Reverse Transcription Polymerase Chain Reaction, Expressing, Western Blot

    Overexpression of IDO affected the MAPK/ERK pathway. Western blot assay was performed on the protein expression levels of (A) p-Raf-1, Raf-1, p-Mek1/2 Mek1/2 (B) p-Erk1/2 and (C) Erk1/2 in 16HBE cells treated with control, NC, DEX, NC+DEX, and IDO+DEX. *P

    Journal: Experimental and Therapeutic Medicine

    Article Title: Overexpression of indoleamine 2, 3-dioxygenase contributes to the repair of human airway epithelial cells inhibited by dexamethasone via affecting the MAPK/ERK signaling pathway

    doi: 10.3892/etm.2018.6163

    Figure Lengend Snippet: Overexpression of IDO affected the MAPK/ERK pathway. Western blot assay was performed on the protein expression levels of (A) p-Raf-1, Raf-1, p-Mek1/2 Mek1/2 (B) p-Erk1/2 and (C) Erk1/2 in 16HBE cells treated with control, NC, DEX, NC+DEX, and IDO+DEX. *P

    Article Snippet: After blocking, the membranes were incubated with anti-IDO (dilution, 1:500; ab55305); anti-p-Raf-1 (dilution, 1:500; ab208449); anti-Raf-1 (dilution, 1:1,000; ab50858); anti-p-Mek1/2 (dilution, 1:1,000; ab194754); anti-Mek1/2 (dilution, 1:1,000; ab215263); anti-p-Erk1/2 (dilution, 1:1,000; ab201015); anti-Erk1/2 (dilution, 1:1,000; ab17942); anti-GAPDH (dilution, 1:1,000, ab8245; all from Abcam) antibodies overnight at 4°C.

    Techniques: Over Expression, Western Blot, Expressing

    Lidocaine inhibited MEK/ERK and NF-κB pathways by up-regulating miR-145. a The phosphorylation of MEK/ERK and ( b ) p65 and IκBα were decreased by lidocaine. The phosphorylation of MEK/ERK, p65 and IκBα were analyzed by western blot. ** p

    Journal: BMC Cancer

    Article Title: Lidocaine inhibits growth, migration and invasion of gastric carcinoma cells by up-regulation of miR-145

    doi: 10.1186/s12885-019-5431-9

    Figure Lengend Snippet: Lidocaine inhibited MEK/ERK and NF-κB pathways by up-regulating miR-145. a The phosphorylation of MEK/ERK and ( b ) p65 and IκBα were decreased by lidocaine. The phosphorylation of MEK/ERK, p65 and IκBα were analyzed by western blot. ** p

    Article Snippet: The membranes were incubated with the primary antibodies against Bcl-2 (ab32124), cleaved-Casapse-3 (ab49822), cleaved-Caspase-7 (ab32522), cleaved-Caspase-9 (ab52298), MMP-2 (ab37150), MMP-9 (ab73734), Vimentin (ab8978), MEK (ab32576), p-MEK (ab96379), ERK (ab32537), p-ERK (ab131438), p65 (ab16502), p-p65 (ab86299), IκBα (ab32518), p-IκBα (ab32518), and β-actin (ab8227) purchased from Abcam (Cambridge, UK) at the dilution of 1:1000.

    Techniques: Western Blot

    MEK1 is required to inhibit the expression of the pluripotency genes pou5f3.2 and ventx2 . ( A ) Embryos injected with 25 ng Mk-MO at 16-cell stage in one animal dorsal blastomere were grown until late gastrulation stage 13 and processed for WISH with pou5f3.2 and ventx2 probes. ( B ) Embryos injected with 25 ng Mk-MO at 16 cell stage in one animal ventral blastomere were grown until mid-neurula stage 18 and processed for WISH with pou5f3.2 probe. ( C ) Four-cell embryos were injected in each blastomere with 25 ng Mk-MO and grown until blastula stage 9, when animal caps were isolated, cultured in vitro until late gastrula stage 13 and then processed for RT-qPCR. In A and B, the number of embryos exemplified by the photograph over the total number of embryos analyzed is indicated. DOI: http://dx.doi.org/10.7554/eLife.21526.008

    Journal: eLife

    Article Title: Lineage commitment of embryonic cells involves MEK1-dependent clearance of pluripotency regulator Ventx2

    doi: 10.7554/eLife.21526

    Figure Lengend Snippet: MEK1 is required to inhibit the expression of the pluripotency genes pou5f3.2 and ventx2 . ( A ) Embryos injected with 25 ng Mk-MO at 16-cell stage in one animal dorsal blastomere were grown until late gastrulation stage 13 and processed for WISH with pou5f3.2 and ventx2 probes. ( B ) Embryos injected with 25 ng Mk-MO at 16 cell stage in one animal ventral blastomere were grown until mid-neurula stage 18 and processed for WISH with pou5f3.2 probe. ( C ) Four-cell embryos were injected in each blastomere with 25 ng Mk-MO and grown until blastula stage 9, when animal caps were isolated, cultured in vitro until late gastrula stage 13 and then processed for RT-qPCR. In A and B, the number of embryos exemplified by the photograph over the total number of embryos analyzed is indicated. DOI: http://dx.doi.org/10.7554/eLife.21526.008

    Article Snippet: Primary antibodies were as follows: anti-Myc (9E10; Santa Cruz Biotech, dilution 1/300 RRID: AB_627268 ), anti-GFP (GFP-1020; 2BScientific; dilution 1/1000 RRID: AB_10000240 ), anti-γ-Tubulin (ab16504; Abcam; dilution 1/1000 RRID: AB_443396 ), anti-phospho-MEK1 (9121; Cell Signaling Technology; dilution 1/400 RRID: AB_331648 ).

    Techniques: Expressing, Injection, Isolation, Cell Culture, In Vitro, Quantitative RT-PCR

    Neural induction in vivo depends on MEK1 activity. Sixteen-cell embryos were injected in one ventral-animal blastomere with 3 ng of dominant-negative Smad5 (Smad5sbn) mRNA and 25 ng Mk-MO ATG, as indicated. Embryos were fixed at late gastrula stage 13, and processed for WISH with the indicated probes. The number of embryos exemplified by the photograph over the total number of embryos analyzed is indicated. Lateral views. DOI: http://dx.doi.org/10.7554/eLife.21526.007

    Journal: eLife

    Article Title: Lineage commitment of embryonic cells involves MEK1-dependent clearance of pluripotency regulator Ventx2

    doi: 10.7554/eLife.21526

    Figure Lengend Snippet: Neural induction in vivo depends on MEK1 activity. Sixteen-cell embryos were injected in one ventral-animal blastomere with 3 ng of dominant-negative Smad5 (Smad5sbn) mRNA and 25 ng Mk-MO ATG, as indicated. Embryos were fixed at late gastrula stage 13, and processed for WISH with the indicated probes. The number of embryos exemplified by the photograph over the total number of embryos analyzed is indicated. Lateral views. DOI: http://dx.doi.org/10.7554/eLife.21526.007

    Article Snippet: Primary antibodies were as follows: anti-Myc (9E10; Santa Cruz Biotech, dilution 1/300 RRID: AB_627268 ), anti-GFP (GFP-1020; 2BScientific; dilution 1/1000 RRID: AB_10000240 ), anti-γ-Tubulin (ab16504; Abcam; dilution 1/1000 RRID: AB_443396 ), anti-phospho-MEK1 (9121; Cell Signaling Technology; dilution 1/400 RRID: AB_331648 ).

    Techniques: In Vivo, Activity Assay, Injection, Dominant Negative Mutation

    Ventx2 degradation and asymmetric distribution require MEK1 activity. ( A ) In silico analysis of phosphorylation sites in the Ventx2 protein and prediction of kinases involved, with Kinasephos2 software. ( B ) Schematic representation of the Ventx2 protein. HD indicates the homeodomain (blue box), and the PEST destruction motif is highlighted in red. Note that Serine 140, which is required for Ventx2 degradation, is a predicted target of MAPK. ( C ) 50 pg Ventx2-Myc RNA was injected into both blastomeres at the two-cell stage. 50 pg GFP-Myc-RNA was co-injected as an internal loading control. Embryos were allowed to develop until the indicated stages, and exogenous Ventx2 was detected by anti-Myc immunostaining on Western blot. The graph shows the ratios of Ventx2-Myc over a-tub signals measured from the Western blot. ( D ) Four-cell embryos were injected in each cell with 50 pg GFP-CAAX, 50 pg Ventx2-Myc, 50 pg 2SAVentx2-Myc RNAs and with 25 ng Mk-MO as indicated. Embryos were fixed at blastula stage 9, cryosectioned and processed for anti-Myc (red), anti-GFP (green) and anti-g-tubulin (centrosomes, white) immunostaining, and DNA was stained with DAPI (blue). Panels represent compiled confocal slices to visualize entire mitotic nuclei. DOI: http://dx.doi.org/10.7554/eLife.21526.011

    Journal: eLife

    Article Title: Lineage commitment of embryonic cells involves MEK1-dependent clearance of pluripotency regulator Ventx2

    doi: 10.7554/eLife.21526

    Figure Lengend Snippet: Ventx2 degradation and asymmetric distribution require MEK1 activity. ( A ) In silico analysis of phosphorylation sites in the Ventx2 protein and prediction of kinases involved, with Kinasephos2 software. ( B ) Schematic representation of the Ventx2 protein. HD indicates the homeodomain (blue box), and the PEST destruction motif is highlighted in red. Note that Serine 140, which is required for Ventx2 degradation, is a predicted target of MAPK. ( C ) 50 pg Ventx2-Myc RNA was injected into both blastomeres at the two-cell stage. 50 pg GFP-Myc-RNA was co-injected as an internal loading control. Embryos were allowed to develop until the indicated stages, and exogenous Ventx2 was detected by anti-Myc immunostaining on Western blot. The graph shows the ratios of Ventx2-Myc over a-tub signals measured from the Western blot. ( D ) Four-cell embryos were injected in each cell with 50 pg GFP-CAAX, 50 pg Ventx2-Myc, 50 pg 2SAVentx2-Myc RNAs and with 25 ng Mk-MO as indicated. Embryos were fixed at blastula stage 9, cryosectioned and processed for anti-Myc (red), anti-GFP (green) and anti-g-tubulin (centrosomes, white) immunostaining, and DNA was stained with DAPI (blue). Panels represent compiled confocal slices to visualize entire mitotic nuclei. DOI: http://dx.doi.org/10.7554/eLife.21526.011

    Article Snippet: Primary antibodies were as follows: anti-Myc (9E10; Santa Cruz Biotech, dilution 1/300 RRID: AB_627268 ), anti-GFP (GFP-1020; 2BScientific; dilution 1/1000 RRID: AB_10000240 ), anti-γ-Tubulin (ab16504; Abcam; dilution 1/1000 RRID: AB_443396 ), anti-phospho-MEK1 (9121; Cell Signaling Technology; dilution 1/400 RRID: AB_331648 ).

    Techniques: Activity Assay, In Silico, Software, Injection, Immunostaining, Western Blot, Staining

    Ventx2 knockdown restores germ-layer formation in MEK1-deficient embryos. ( A ) Four-cell embryos were injected with 50 pg 2SAVentx2-Myc RNA per cell, fixed at tailbud stage 25 and processed for WISH with pou5f3.2 probe. ( B ) Four-cell embryos were injected with 30 ng Ventx2-MO (Vx2-MO) per blastomere, collected at stage 10.5 and processed for RT-qPCR. ( C ) Embryos injected as in B were processed for WISH analysis at early gastrula stage 10.5 with ventx1 and gsc probes (vegetal view). ( D ). Four-cell embryos were injected with 25 ng Mk-MO, with or without 7.5 ng Vx2-MO, in each blastomere, collected at gastrula stage 10.5, and processed for WISH with indicated probes. Note that embryos stained for xk81a1 (epidermis) were injected in one ventral animal blastomere at 16 cell stage and collected at late gastrula stage 13. Embryos stained for gsc were hemisectioned prior to staining to improve probe penetration. In A and D, the number of embryos exemplified by the photograph over the total number of embryos analyzed is indicated. For the RT-qPCR graph, error bars represent s.e.m. values of three independent experiments with two technical duplicates. For statistical analysis, samples from injected embryos were compared with samples from uninjected control embryos by Unpaired Student’s t-test. *p

    Journal: eLife

    Article Title: Lineage commitment of embryonic cells involves MEK1-dependent clearance of pluripotency regulator Ventx2

    doi: 10.7554/eLife.21526

    Figure Lengend Snippet: Ventx2 knockdown restores germ-layer formation in MEK1-deficient embryos. ( A ) Four-cell embryos were injected with 50 pg 2SAVentx2-Myc RNA per cell, fixed at tailbud stage 25 and processed for WISH with pou5f3.2 probe. ( B ) Four-cell embryos were injected with 30 ng Ventx2-MO (Vx2-MO) per blastomere, collected at stage 10.5 and processed for RT-qPCR. ( C ) Embryos injected as in B were processed for WISH analysis at early gastrula stage 10.5 with ventx1 and gsc probes (vegetal view). ( D ). Four-cell embryos were injected with 25 ng Mk-MO, with or without 7.5 ng Vx2-MO, in each blastomere, collected at gastrula stage 10.5, and processed for WISH with indicated probes. Note that embryos stained for xk81a1 (epidermis) were injected in one ventral animal blastomere at 16 cell stage and collected at late gastrula stage 13. Embryos stained for gsc were hemisectioned prior to staining to improve probe penetration. In A and D, the number of embryos exemplified by the photograph over the total number of embryos analyzed is indicated. For the RT-qPCR graph, error bars represent s.e.m. values of three independent experiments with two technical duplicates. For statistical analysis, samples from injected embryos were compared with samples from uninjected control embryos by Unpaired Student’s t-test. *p

    Article Snippet: Primary antibodies were as follows: anti-Myc (9E10; Santa Cruz Biotech, dilution 1/300 RRID: AB_627268 ), anti-GFP (GFP-1020; 2BScientific; dilution 1/1000 RRID: AB_10000240 ), anti-γ-Tubulin (ab16504; Abcam; dilution 1/1000 RRID: AB_443396 ), anti-phospho-MEK1 (9121; Cell Signaling Technology; dilution 1/400 RRID: AB_331648 ).

    Techniques: Injection, Quantitative RT-PCR, Staining

    MEK1 depletion by morpholinos. ( A ) Four-cell embryos were injected in each blastomere with 50 pg GFP‐CAAX mRNA with or without 25 ng Mk-MO, fixed at blastula stage 9, cryosectioned and stained with anti-phospho-MEK1 antibody. The pMEK1 signal was severely reduced or lost in cells injected with Mk-MO. ( B ) Four-cell embryos were injected in each cell with 25 ng Mk-MO, collected at early gastrula stage 10.5 and processed for RT-qPCR to quantify p53 expression levels. Mk‐MO did not induce p53 expression. ( C ) Two-cell embryos were injected twice with 25 ng Mk-MO in each blastomere, followed by injection at 4 cell stage of hamster MEK1 mRNA (Mk; 400 pg per blastomere). In order to score progress through gastrulation pictures were taken from live stage 13 gastrula embryos (vegetal view). ( D ) Blastopore closure was scored by calculating the ratio of blastopore diameter of injected embryos to the mean of blastopore diameter of uninjected control embryos. Bars represent maximum and minimum values, and lines represent the mean. The number of embryos analyzed in each condition is displayed on the graph. For statistical analysis, samples were compared by Mann-Whitney test (99% confidence intervals were applied; ***p≤0.0001). DOI: http://dx.doi.org/10.7554/eLife.21526.003 10.7554/eLife.21526.004 Values of blastopore closure ratios. Details are shown in Figure 1—figure supplement 1 and Materials and methods. DOI: http://dx.doi.org/10.7554/eLife.21526.004

    Journal: eLife

    Article Title: Lineage commitment of embryonic cells involves MEK1-dependent clearance of pluripotency regulator Ventx2

    doi: 10.7554/eLife.21526

    Figure Lengend Snippet: MEK1 depletion by morpholinos. ( A ) Four-cell embryos were injected in each blastomere with 50 pg GFP‐CAAX mRNA with or without 25 ng Mk-MO, fixed at blastula stage 9, cryosectioned and stained with anti-phospho-MEK1 antibody. The pMEK1 signal was severely reduced or lost in cells injected with Mk-MO. ( B ) Four-cell embryos were injected in each cell with 25 ng Mk-MO, collected at early gastrula stage 10.5 and processed for RT-qPCR to quantify p53 expression levels. Mk‐MO did not induce p53 expression. ( C ) Two-cell embryos were injected twice with 25 ng Mk-MO in each blastomere, followed by injection at 4 cell stage of hamster MEK1 mRNA (Mk; 400 pg per blastomere). In order to score progress through gastrulation pictures were taken from live stage 13 gastrula embryos (vegetal view). ( D ) Blastopore closure was scored by calculating the ratio of blastopore diameter of injected embryos to the mean of blastopore diameter of uninjected control embryos. Bars represent maximum and minimum values, and lines represent the mean. The number of embryos analyzed in each condition is displayed on the graph. For statistical analysis, samples were compared by Mann-Whitney test (99% confidence intervals were applied; ***p≤0.0001). DOI: http://dx.doi.org/10.7554/eLife.21526.003 10.7554/eLife.21526.004 Values of blastopore closure ratios. Details are shown in Figure 1—figure supplement 1 and Materials and methods. DOI: http://dx.doi.org/10.7554/eLife.21526.004

    Article Snippet: Primary antibodies were as follows: anti-Myc (9E10; Santa Cruz Biotech, dilution 1/300 RRID: AB_627268 ), anti-GFP (GFP-1020; 2BScientific; dilution 1/1000 RRID: AB_10000240 ), anti-γ-Tubulin (ab16504; Abcam; dilution 1/1000 RRID: AB_443396 ), anti-phospho-MEK1 (9121; Cell Signaling Technology; dilution 1/400 RRID: AB_331648 ).

    Techniques: Injection, Staining, Quantitative RT-PCR, Expressing, MANN-WHITNEY

    Gene expression analysis of MEK1-depleted gastrula embryos. ( A–D ) Four-cell embryos were injected in each blastomere with 25 ng Mk-MO, collected at early gastrula stage 10.5 and processed for RT-qPCR to quantify changes in the expression levels of pro-differentiation markers ( A ), or changes in the expression levels of BMPs (Smad1/5/8) and Activin/Nodal (Smad2/3) signaling targets ( C ). For all qPCR graphs, error bars represent s.e.m. values of four independent experiments with two technical duplicates. For statistical analyses, samples from injected embryos were compared with samples from uninjected control embryos by Unpaired Student’s t-test. *p

    Journal: eLife

    Article Title: Lineage commitment of embryonic cells involves MEK1-dependent clearance of pluripotency regulator Ventx2

    doi: 10.7554/eLife.21526

    Figure Lengend Snippet: Gene expression analysis of MEK1-depleted gastrula embryos. ( A–D ) Four-cell embryos were injected in each blastomere with 25 ng Mk-MO, collected at early gastrula stage 10.5 and processed for RT-qPCR to quantify changes in the expression levels of pro-differentiation markers ( A ), or changes in the expression levels of BMPs (Smad1/5/8) and Activin/Nodal (Smad2/3) signaling targets ( C ). For all qPCR graphs, error bars represent s.e.m. values of four independent experiments with two technical duplicates. For statistical analyses, samples from injected embryos were compared with samples from uninjected control embryos by Unpaired Student’s t-test. *p

    Article Snippet: Primary antibodies were as follows: anti-Myc (9E10; Santa Cruz Biotech, dilution 1/300 RRID: AB_627268 ), anti-GFP (GFP-1020; 2BScientific; dilution 1/1000 RRID: AB_10000240 ), anti-γ-Tubulin (ab16504; Abcam; dilution 1/1000 RRID: AB_443396 ), anti-phospho-MEK1 (9121; Cell Signaling Technology; dilution 1/400 RRID: AB_331648 ).

    Techniques: Expressing, Injection, Quantitative RT-PCR, Real-time Polymerase Chain Reaction