Aberrant neuronal activity-induced signaling and gene expression in a mouse model of RASopathy
Fig 5
Neuronal activity-induced phosphorylation of ERK is disturbed in Ptpn11D61Y.
(A) Exemplary photograph of an acute slice used for experiments. (B) Western blot of lysates from control and Ptpn11D61Y acute slices treated with 4AP/Bic (+) or vehicle (-) for 10 minutes and probed with antibodies against pERK, ERK and β-III-tubulin. The latter was used as a loading control. (C-E) Quantification of the Western blot experiment as exemplified in B is shown. The stimulation of control slices led to a significant increase of the pERK level (C, D). The basal pERK level was increased in Ptpn11D61Y slices compared to controls. No further increase of pERK immunoreactivity could be detected upon stimulation of neuronal activity (C, D). Note the increased total expression of ERK in the slices from Ptpn11D61Y in the basal state and upon stimulation (E). (F-H) The quantification of ERK phosphorylation in the nuclear fraction prepared from forebrains indicates an increase in the pERK level in the samples from Ptpn11D61Y animals as compared to controls. Data are shown as mean ± SEM and analyzed using either one-way ANOVA followed by Bonferroni´s multiple comparison test or unpaired t-test (*p≤0.05, ***p≤0.001). The number of replicates from a total of three independent experiments is indicated in the columns of the graph.
