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Acute 6-OH–BDE-47 exposure impairs <t>MEK–ERK</t> signaling. A, top panel: synaptosomes were prepared from neurons acutely exposed (20 min pre-treatment) to 5 μm 6-OH with and without Bic + 4AP stimulus (10 min). Levels of the CaMK1 target pSynapsin1(serine 9) were assessed via Western blotting. n = 2. Bottom panel: to reproduce a previously reported effect linking CaMK1 to MEK–ERK activity (70), neurons were acutely exposed (60 min pre-treatment) to the CaMKK inhibitor STO-609 prior to treatment with Bic + 4AP for 10 min (indicated as +B10′). Inhibition of pERK induction was assessed by Western blotting and is quantified below. n = 3, p value generated by unpaired one-tailed t test. B, cells acutely exposed (20 min pre-treatment) to BDE-47 or one of its hydroxylated metabolites were stimulated with Bic + 4AP for 10 min before collecting whole-cell lysates. Activation of MEK–ERK signaling was assessed by Western blotting, measuring pERK <t>and</t> <t>pMEK</t> levels. Note, the lower band in the MEK panel is the ERK band visualized on the same blot. Quantification relative to total ERK and MEK is shown below representative blots. n = 3–6, p values generated by one-way ANOVA with post hoc LSD. pERK quantification: treatment F(7,22) = 9.822, p < 0.0001. pMEK quantification: treatment F(7,16) = 7.805, p = 0.0004. C, same experimental design as in B., but neurons were stimulated with TTX + PMA to induce MEK–ERK signaling synapse-independently. n = 3. pERK quantification: treatment F(7,16) = 37.95, p < 0.0001.
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Acute 6-OH–BDE-47 exposure impairs <t>MEK–ERK</t> signaling. A, top panel: synaptosomes were prepared from neurons acutely exposed (20 min pre-treatment) to 5 μm 6-OH with and without Bic + 4AP stimulus (10 min). Levels of the CaMK1 target pSynapsin1(serine 9) were assessed via Western blotting. n = 2. Bottom panel: to reproduce a previously reported effect linking CaMK1 to MEK–ERK activity (70), neurons were acutely exposed (60 min pre-treatment) to the CaMKK inhibitor STO-609 prior to treatment with Bic + 4AP for 10 min (indicated as +B10′). Inhibition of pERK induction was assessed by Western blotting and is quantified below. n = 3, p value generated by unpaired one-tailed t test. B, cells acutely exposed (20 min pre-treatment) to BDE-47 or one of its hydroxylated metabolites were stimulated with Bic + 4AP for 10 min before collecting whole-cell lysates. Activation of MEK–ERK signaling was assessed by Western blotting, measuring pERK <t>and</t> <t>pMEK</t> levels. Note, the lower band in the MEK panel is the ERK band visualized on the same blot. Quantification relative to total ERK and MEK is shown below representative blots. n = 3–6, p values generated by one-way ANOVA with post hoc LSD. pERK quantification: treatment F(7,22) = 9.822, p < 0.0001. pMEK quantification: treatment F(7,16) = 7.805, p = 0.0004. C, same experimental design as in B., but neurons were stimulated with TTX + PMA to induce MEK–ERK signaling synapse-independently. n = 3. pERK quantification: treatment F(7,16) = 37.95, p < 0.0001.
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Acute 6-OH–BDE-47 exposure impairs <t>MEK–ERK</t> signaling. A, top panel: synaptosomes were prepared from neurons acutely exposed (20 min pre-treatment) to 5 μm 6-OH with and without Bic + 4AP stimulus (10 min). Levels of the CaMK1 target pSynapsin1(serine 9) were assessed via Western blotting. n = 2. Bottom panel: to reproduce a previously reported effect linking CaMK1 to MEK–ERK activity (70), neurons were acutely exposed (60 min pre-treatment) to the CaMKK inhibitor STO-609 prior to treatment with Bic + 4AP for 10 min (indicated as +B10′). Inhibition of pERK induction was assessed by Western blotting and is quantified below. n = 3, p value generated by unpaired one-tailed t test. B, cells acutely exposed (20 min pre-treatment) to BDE-47 or one of its hydroxylated metabolites were stimulated with Bic + 4AP for 10 min before collecting whole-cell lysates. Activation of MEK–ERK signaling was assessed by Western blotting, measuring pERK <t>and</t> <t>pMEK</t> levels. Note, the lower band in the MEK panel is the ERK band visualized on the same blot. Quantification relative to total ERK and MEK is shown below representative blots. n = 3–6, p values generated by one-way ANOVA with post hoc LSD. pERK quantification: treatment F(7,22) = 9.822, p < 0.0001. pMEK quantification: treatment F(7,16) = 7.805, p = 0.0004. C, same experimental design as in B., but neurons were stimulated with TTX + PMA to induce MEK–ERK signaling synapse-independently. n = 3. pERK quantification: treatment F(7,16) = 37.95, p < 0.0001.
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anti mek  (Cell Signaling Technology Inc)
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Acute 6-OH–BDE-47 exposure impairs <t>MEK–ERK</t> signaling. A, top panel: synaptosomes were prepared from neurons acutely exposed (20 min pre-treatment) to 5 μm 6-OH with and without Bic + 4AP stimulus (10 min). Levels of the CaMK1 target pSynapsin1(serine 9) were assessed via Western blotting. n = 2. Bottom panel: to reproduce a previously reported effect linking CaMK1 to MEK–ERK activity (70), neurons were acutely exposed (60 min pre-treatment) to the CaMKK inhibitor STO-609 prior to treatment with Bic + 4AP for 10 min (indicated as +B10′). Inhibition of pERK induction was assessed by Western blotting and is quantified below. n = 3, p value generated by unpaired one-tailed t test. B, cells acutely exposed (20 min pre-treatment) to BDE-47 or one of its hydroxylated metabolites were stimulated with Bic + 4AP for 10 min before collecting whole-cell lysates. Activation of MEK–ERK signaling was assessed by Western blotting, measuring pERK <t>and</t> <t>pMEK</t> levels. Note, the lower band in the MEK panel is the ERK band visualized on the same blot. Quantification relative to total ERK and MEK is shown below representative blots. n = 3–6, p values generated by one-way ANOVA with post hoc LSD. pERK quantification: treatment F(7,22) = 9.822, p < 0.0001. pMEK quantification: treatment F(7,16) = 7.805, p = 0.0004. C, same experimental design as in B., but neurons were stimulated with TTX + PMA to induce MEK–ERK signaling synapse-independently. n = 3. pERK quantification: treatment F(7,16) = 37.95, p < 0.0001.
Anti Mek, supplied by Cell Signaling Technology Inc, 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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Acute 6-OH–BDE-47 exposure impairs <t>MEK–ERK</t> signaling. A, top panel: synaptosomes were prepared from neurons acutely exposed (20 min pre-treatment) to 5 μm 6-OH with and without Bic + 4AP stimulus (10 min). Levels of the CaMK1 target pSynapsin1(serine 9) were assessed via Western blotting. n = 2. Bottom panel: to reproduce a previously reported effect linking CaMK1 to MEK–ERK activity (70), neurons were acutely exposed (60 min pre-treatment) to the CaMKK inhibitor STO-609 prior to treatment with Bic + 4AP for 10 min (indicated as +B10′). Inhibition of pERK induction was assessed by Western blotting and is quantified below. n = 3, p value generated by unpaired one-tailed t test. B, cells acutely exposed (20 min pre-treatment) to BDE-47 or one of its hydroxylated metabolites were stimulated with Bic + 4AP for 10 min before collecting whole-cell lysates. Activation of MEK–ERK signaling was assessed by Western blotting, measuring pERK <t>and</t> <t>pMEK</t> levels. Note, the lower band in the MEK panel is the ERK band visualized on the same blot. Quantification relative to total ERK and MEK is shown below representative blots. n = 3–6, p values generated by one-way ANOVA with post hoc LSD. pERK quantification: treatment F(7,22) = 9.822, p < 0.0001. pMEK quantification: treatment F(7,16) = 7.805, p = 0.0004. C, same experimental design as in B., but neurons were stimulated with TTX + PMA to induce MEK–ERK signaling synapse-independently. n = 3. pERK quantification: treatment F(7,16) = 37.95, p < 0.0001.
Anti Mek, supplied by Cell Signaling Technology Inc, 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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Acute 6-OH–BDE-47 exposure impairs <t>MEK–ERK</t> signaling. A, top panel: synaptosomes were prepared from neurons acutely exposed (20 min pre-treatment) to 5 μm 6-OH with and without Bic + 4AP stimulus (10 min). Levels of the CaMK1 target pSynapsin1(serine 9) were assessed via Western blotting. n = 2. Bottom panel: to reproduce a previously reported effect linking CaMK1 to MEK–ERK activity (70), neurons were acutely exposed (60 min pre-treatment) to the CaMKK inhibitor STO-609 prior to treatment with Bic + 4AP for 10 min (indicated as +B10′). Inhibition of pERK induction was assessed by Western blotting and is quantified below. n = 3, p value generated by unpaired one-tailed t test. B, cells acutely exposed (20 min pre-treatment) to BDE-47 or one of its hydroxylated metabolites were stimulated with Bic + 4AP for 10 min before collecting whole-cell lysates. Activation of MEK–ERK signaling was assessed by Western blotting, measuring pERK <t>and</t> <t>pMEK</t> levels. Note, the lower band in the MEK panel is the ERK band visualized on the same blot. Quantification relative to total ERK and MEK is shown below representative blots. n = 3–6, p values generated by one-way ANOVA with post hoc LSD. pERK quantification: treatment F(7,22) = 9.822, p < 0.0001. pMEK quantification: treatment F(7,16) = 7.805, p = 0.0004. C, same experimental design as in B., but neurons were stimulated with TTX + PMA to induce MEK–ERK signaling synapse-independently. n = 3. pERK quantification: treatment F(7,16) = 37.95, p < 0.0001.
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Image Search Results


Acute 6-OH–BDE-47 exposure impairs MEK–ERK signaling. A, top panel: synaptosomes were prepared from neurons acutely exposed (20 min pre-treatment) to 5 μm 6-OH with and without Bic + 4AP stimulus (10 min). Levels of the CaMK1 target pSynapsin1(serine 9) were assessed via Western blotting. n = 2. Bottom panel: to reproduce a previously reported effect linking CaMK1 to MEK–ERK activity (70), neurons were acutely exposed (60 min pre-treatment) to the CaMKK inhibitor STO-609 prior to treatment with Bic + 4AP for 10 min (indicated as +B10′). Inhibition of pERK induction was assessed by Western blotting and is quantified below. n = 3, p value generated by unpaired one-tailed t test. B, cells acutely exposed (20 min pre-treatment) to BDE-47 or one of its hydroxylated metabolites were stimulated with Bic + 4AP for 10 min before collecting whole-cell lysates. Activation of MEK–ERK signaling was assessed by Western blotting, measuring pERK and pMEK levels. Note, the lower band in the MEK panel is the ERK band visualized on the same blot. Quantification relative to total ERK and MEK is shown below representative blots. n = 3–6, p values generated by one-way ANOVA with post hoc LSD. pERK quantification: treatment F(7,22) = 9.822, p < 0.0001. pMEK quantification: treatment F(7,16) = 7.805, p = 0.0004. C, same experimental design as in B., but neurons were stimulated with TTX + PMA to induce MEK–ERK signaling synapse-independently. n = 3. pERK quantification: treatment F(7,16) = 37.95, p < 0.0001.

Journal: The Journal of Biological Chemistry

Article Title: Certain ortho-hydroxylated brominated ethers are promiscuous kinase inhibitors that impair neuronal signaling and neurodevelopmental processes

doi: 10.1074/jbc.RA119.011138

Figure Lengend Snippet: Acute 6-OH–BDE-47 exposure impairs MEK–ERK signaling. A, top panel: synaptosomes were prepared from neurons acutely exposed (20 min pre-treatment) to 5 μm 6-OH with and without Bic + 4AP stimulus (10 min). Levels of the CaMK1 target pSynapsin1(serine 9) were assessed via Western blotting. n = 2. Bottom panel: to reproduce a previously reported effect linking CaMK1 to MEK–ERK activity (70), neurons were acutely exposed (60 min pre-treatment) to the CaMKK inhibitor STO-609 prior to treatment with Bic + 4AP for 10 min (indicated as +B10′). Inhibition of pERK induction was assessed by Western blotting and is quantified below. n = 3, p value generated by unpaired one-tailed t test. B, cells acutely exposed (20 min pre-treatment) to BDE-47 or one of its hydroxylated metabolites were stimulated with Bic + 4AP for 10 min before collecting whole-cell lysates. Activation of MEK–ERK signaling was assessed by Western blotting, measuring pERK and pMEK levels. Note, the lower band in the MEK panel is the ERK band visualized on the same blot. Quantification relative to total ERK and MEK is shown below representative blots. n = 3–6, p values generated by one-way ANOVA with post hoc LSD. pERK quantification: treatment F(7,22) = 9.822, p < 0.0001. pMEK quantification: treatment F(7,16) = 7.805, p = 0.0004. C, same experimental design as in B., but neurons were stimulated with TTX + PMA to induce MEK–ERK signaling synapse-independently. n = 3. pERK quantification: treatment F(7,16) = 37.95, p < 0.0001.

Article Snippet: Primary antibodies included the following antibodies: β-actin (Thermo Fisher Scientific, catalog no. AM4302, RRID:AB_2536382 ); H4 (CST catalog no. 2935, RRID:AB_1147658 ); pERK (CST catalog no. 4370, RRID:AB_2315112 ); ERK (CST catalog no. 4696, RRID:AB_390780 ); pMEK (CST catalog no. 9154, RRID:AB_2138017 ); MEK (CST catalog no. 2352, RRID:AB_10693788 ); synapsin1 (SySy catalog no. 106011, RRID:AB_2619772 );PSD-95 (NeuroMab catalog no. 75-028, RRID:AB_2292909 ); gephyrin (SySy catalog no. 147111, RRID:AB_887719 ); synaptotagmin6 (NeuroMab catalog no. 75-271, RRID:AB_11001830 ); and synaptophysin1(SySy catalog no. 101011, RRID:AB_887824 ).

Techniques: Western Blot, Activity Assay, Inhibition, Generated, One-tailed Test, Activation Assay