Journal: The Journal of Neuroscience

Article Title: Shift from Extracellular Signal-Regulated Kinase to AKT/cAMP Response Element-Binding Protein Pathway Increases Survival-Motor-Neuron Expression in Spinal-Muscular-Atrophy-Like Mice and Patient Cells

doi: 10.1523/JNEUROSCI.2728-12.2013

Figure Lengend Snippet: Activated Elk-1 and CREB transcription factors compete for binding the SMN2 promoter in the spinal cord of severe SMA-like mice. A, B, SMN promoter site 1 (A) and site 2 (B) sequences. C–F, Western blot analysis and quantification of Elk-1 protein (C, D) phosphorylation and CREB protein (E, F) phosphorylation in the ventral lumbar spinal cord of vehicle- and U0126-treated control mice at 2 and 6 d of age, vehicle-treated SMA-like mice at 2 d of age, and U0126-treated SMA-like mice at 2 and 6 d of age (n = 3). G–J, ChIP analysis of phospho-Elk-1 (G, H) and phospho-CREB (I, J) in the ventral lumbar spinal cord of vehicle- and U0126-treated control mice at 2 and 6 d of age, vehicle-treated SMA-like mice at 2 d of age, and U0126-treated SMA-like mice at 2 and 6 d of age (n = 9). Quantitative real-time PCR was performed to detect SMN2 promoter site 1 (G, I) and site 2 (H, J). Data are displayed as mean ± SEM. *p < 0.05, **p < 0.01, ***p < 0.001.

Article Snippet: DNA was incubated overnight at 4°C on a rotating wheel with 1 μg of the following antibodies: polyclonal rabbit anti-Ser133 phospho-CREB (Millipore), monoclonal mouse anti-Ser183 phospho-Elk-1 (Santa Cruz Biotechnology), polyclonal rabbit anti-acetyl-histone H3 Lys9 (Millipore), and polyclonal rabbit-acetyl-histone H4 Lys8 (Millipore).

Techniques: Binding Assay, Western Blot, Real-time Polymerase Chain Reaction

Journal: The Journal of Neuroscience

Article Title: Shift from Extracellular Signal-Regulated Kinase to AKT/cAMP Response Element-Binding Protein Pathway Increases Survival-Motor-Neuron Expression in Spinal-Muscular-Atrophy-Like Mice and Patient Cells

doi: 10.1523/JNEUROSCI.2728-12.2013

Figure Lengend Snippet: The crosstalk between ERK and AKT signaling pathways is reciprocal in SMA spinal cord. A, B, Western blot analysis and quantification of ERK protein phosphorylation in control and SMA spinal cord explants in the presence or absence of either NMDA or the CREB inhibitor KG-501 (n = 3). C, D, Western blot analysis and quantification of Elk-1 protein phosphorylation in control and SMA spinal cord explants in the presence or not of NMDA and of the CREB inhibitor KG-501 (n = 3). E, F, Western blot analysis and quantification of AKT protein phosphorylation in control and SMA spinal cord explants in the presence or absence of either NMDA or the CREB inhibitor KG-501 (n = 3). G, H, Western blot analysis and quantification of SMN protein expression in control and SMA spinal cord explants in the presence or absence of NMDA or the CREB inhibitor KG-501 (n = 3). Data are displayed as mean ± SEM. *p < 0.05.

Article Snippet: DNA was incubated overnight at 4°C on a rotating wheel with 1 μg of the following antibodies: polyclonal rabbit anti-Ser133 phospho-CREB (Millipore), monoclonal mouse anti-Ser183 phospho-Elk-1 (Santa Cruz Biotechnology), polyclonal rabbit anti-acetyl-histone H3 Lys9 (Millipore), and polyclonal rabbit-acetyl-histone H4 Lys8 (Millipore).

Techniques: Western Blot, Expressing

Journal: The Journal of Neuroscience

Article Title: Shift from Extracellular Signal-Regulated Kinase to AKT/cAMP Response Element-Binding Protein Pathway Increases Survival-Motor-Neuron Expression in Spinal-Muscular-Atrophy-Like Mice and Patient Cells

doi: 10.1523/JNEUROSCI.2728-12.2013

Figure Lengend Snippet: Proposed mechanisms involved in the increase of SMN expression induced by ERK inhibition in the spinal cord of severe SMA-like mice. A, Activation profile of ERK/Elk-1 and AKT/CREB signaling pathways in lumbar spinal cord of vehicle-treated SMA-like mice compared with control. Large characters indicate activated molecules. B, Activation profile of ERK/Elk-1 and AKT/CREB signaling pathways in lumbar spinal cord of U0126-treated SMA-like mice compared with vehicle-treated SMA-like mice. Large characters indicate activated molecules.

Article Snippet: DNA was incubated overnight at 4°C on a rotating wheel with 1 μg of the following antibodies: polyclonal rabbit anti-Ser133 phospho-CREB (Millipore), monoclonal mouse anti-Ser183 phospho-Elk-1 (Santa Cruz Biotechnology), polyclonal rabbit anti-acetyl-histone H3 Lys9 (Millipore), and polyclonal rabbit-acetyl-histone H4 Lys8 (Millipore).

Techniques: Expressing, Inhibition, Activation Assay