Figure 2: Effects of cycloheximide and rapamycin on BDNF-induced changes in levels of SCOP and p-ERK. | Nature Communications

Figure 2: Effects of cycloheximide and rapamycin on BDNF-induced changes in levels of SCOP and p-ERK.

From: A molecular brake controls the magnitude of long-term potentiation

Figure 2

(a,d) Experimental design: hippocampal slices were treated with BDNF (100 ng ml−1) for the indicated periods of time in the presence of cycloheximide (25 μM) or rapamycin (1 μM). (b,e) Representative western blottings for SCOP, Akt, p-ERK and ERK under various experimental conditions. (c,f) Quantitative analysis of the levels of SCOP (normalized to the values of Akt) and p-ERK/ERK ratios under various experimental conditions. In all cases, results are means±s.e.m. of three experiments. *P<0.05, as compared with time point 0, one-way analysis of variance (ANOVA) followed by Bonferroni test. (g) Representative blot for all newly synthesized (biotin labelled) proteins larger than 150 kDa from synaptoneurosomes before immunoprecipitation (input) as detected by IRDye 800CW Streptavidin. Note that the second lane (BDNF-treated sample) shows denser labelling than the other lanes. NL (not labelled) line is from naive synaptoneurosomes without metabolic labelling (negative control). (h) Representative western blottings for newly synthesized (biotin labelled) and total SCOP, after treatment of cortical synaptoneurosomes with BDNF (100 ng ml−1) in the absence or presence of rapamycin (1 μM), and subsequent immunoprecipitation with SCOP antibody. (i) Quantitative analysis of the levels of newly synthesized SCOP (normalized to total SCOP). Data are means±s.e.m. of four experiments. *P<0.05, as compared with control, two-way ANOVA followed by Bonferroni test.

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