Journal: Journal of Molecular Cell Biology

Article Title: ATP11B deficiency leads to impairment of hippocampal synaptic plasticity

doi: 10.1093/jmcb/mjz042

Figure Lengend Snippet: ATP11B regulates synaptic plasticity through the MAPK14 signaling pathway. ( A ) QPCR detection of the mRNA expression levels of the synaptic proteins Mapk14 , Creb1 , and Arc in primary hippocampal neurons after transfected with overexpression or silencing plasmids of Atp11b . Gapdh was used as the internal control. ( B ) The qPCR products were separated on the agarose gel. ( C ) Western blotting analysis of MAPK14 signaling molecules (p-MAPK14, MAPK14, CREB1, and ARC) in primary hippocampal neurons in the following four groups: Control, ATP11B overexpression (ATP11B), shRNA-Control, and ATP11B silencing (shRNA-ATP11B). ( D ) Data are expressed as a percentage of the control relative to GAPDH after background subtraction. ( E ) p-MAPK14 was measured with a p-MAPK14 antibody and MAPK14 as control. Ratio represents the degree of phosphorylation. Data are expressed as mean ± SEM. One-way ANOVA. * P < 0.05.

Article Snippet: The membranes were blocked with 5% bovine serum albumin–PBS for 1 h, and the following primary antibodies were applied at 4°C overnight: rabbit anti-SV2A (Abcam), rabbit anti-SYP (Boster), rabbit anti-DLG4 ((BBI), rabbit anti-MAPK14 (CST), rabbit anti-pMAPK14 (Abclonal), rabbit anti-CREB1 (BBI), rabbit anti-ARC (BBI), and rabbit anti-ATP11B (Sigma).

Techniques: Expressing, Transfection, Over Expression, Agarose Gel Electrophoresis, Western Blot, shRNA

Journal: Journal of Molecular Cell Biology

Article Title: ATP11B deficiency leads to impairment of hippocampal synaptic plasticity

doi: 10.1093/jmcb/mjz042

Figure Lengend Snippet: SB203580 blocks the regulation of ATP11B on the MAPK14 signaling pathway. ( A ) Western blotting detection of MAPK14 signaling molecules (MAPK14, CREB1, and ARC) in primary hippocampal neurons after pretreatment with SB203580 (20 μ m ) for 1 h and/or transfection with ATP11B. ( B ) Data are expressed as a percentage of the control relative to GAPDH after background subtraction in the following four groups: Control, treatment with SB203580 (SB203580), transfection with ATP11B (ATP11B), and transfection with ATP11B following pretreatment with SB203580 (SB203580 + ATP11B). Data are expressed as mean ± SEM. One-way ANOVA. * P < 0.05, *** P < 0.001.

Article Snippet: The membranes were blocked with 5% bovine serum albumin–PBS for 1 h, and the following primary antibodies were applied at 4°C overnight: rabbit anti-SV2A (Abcam), rabbit anti-SYP (Boster), rabbit anti-DLG4 ((BBI), rabbit anti-MAPK14 (CST), rabbit anti-pMAPK14 (Abclonal), rabbit anti-CREB1 (BBI), rabbit anti-ARC (BBI), and rabbit anti-ATP11B (Sigma).

Techniques: Western Blot, Transfection

Journal: Journal of Molecular Cell Biology

Article Title: ATP11B deficiency leads to impairment of hippocampal synaptic plasticity

doi: 10.1093/jmcb/mjz042

Figure Lengend Snippet: A list of primers used for quantitative real-time PCR.

Article Snippet: The membranes were blocked with 5% bovine serum albumin–PBS for 1 h, and the following primary antibodies were applied at 4°C overnight: rabbit anti-SV2A (Abcam), rabbit anti-SYP (Boster), rabbit anti-DLG4 ((BBI), rabbit anti-MAPK14 (CST), rabbit anti-pMAPK14 (Abclonal), rabbit anti-CREB1 (BBI), rabbit anti-ARC (BBI), and rabbit anti-ATP11B (Sigma).

Techniques: