Journal: The EMBO Journal

Article Title: Different Ras isoforms regulate synaptic plasticity in opposite directions

doi: 10.1038/s44318-025-00390-8

Figure Lengend Snippet: ( A ) Quantification (upper panel) and representative Western blot (lower panel) of active Ras pulled-down vs total Ras from hippocampal slice cultures after inducing different chemical plasticity protocols. Both Ras bands were quantified together and results were normalized to control, non-stimulated slices (dashed line) and expressed as mean ± SEM, n = 5–13 independent slice batches; individual values are also represented. One-way ANOVA [F(5,42) = 23.7, p < 0.0001] + Dunnett’s post-test (*) was used to evaluate Ras activation compared to control. ( B ) Time course of Ras activity upon DHPG stimulation in spines (blue, n = 50) and dendrites (orange, n = 15) of CA1 pyramidal cells ( n = 10) evaluated by FRET. Results are represented as mean ± SEM. Representative images of the FRET signal from dendritic branches of CA1 neurons coexpressing H-Ras-GFP and FRas2-M obtained during the time course are shown. Scale bars 2 μm. White arrows point dendritic spines. Kruskal–Wallis [H(11,571) = 53.8, p < 0.0001 for spines, and H(11,178) = 14.1, p = 0.2 for dendrites] + Dunn’s post-test (*) was used to evaluate differences across time (comparing to baseline). ( C ) Quantification (upper panel) and representative Western blot (lower panel) of phospho-Erk (pErk) and Erk signals from hippocampal slices treated with vehicle (DMSO) or inhibitors of CaMKII (KN93), Src family (PP2) or PKC (GF109203) in response to DHPG stimulation (green columns). The phospho-to-total ratio was calculated and normalized to DMSO control condition. Results are expressed as mean ± SEM and individual values are also represented, n = 4–8 independent slice batches. Mixed effects analysis [F(1,7) = 30.0, p = 0.0009] + Sidak’s post-test (*) was used to evaluate DHPG effect for each condition. ( D ) Quantification (upper panel) and representative Western blot (lower panel) of active Ras pulled-down vs total Ras from hippocampal slice cultures treated with vehicle DMSO or PP2 and stimulated with DHPG. Results are normalized to vehicle non-stimulated slices and expressed as mean ± SEM, n = 4 independent slice batches; individual values are also represented. Mixed effects analysis [F(1,6) = 18.6, p = 0.005] + Sidak’s post-test (*) was used to evaluate DHPG effect for each condition. ( E ) Time course (left) of normalized AMPAR-mediated EPSCs from baseline to 40 min after mGluR-LTD induction (PP-LFS, 1 Hz) from organotypic slices treated with DMSO (gray), Src family inhibitor PP2 (blue) or c-Src inhibitor KBSRC4 (pink). Representative traces are shown for baseline responses (dashed line) and for the last 10 min of the recording (thick line). Quantification (right) of the extent of depression 30–40 min after induction. Results are normalized to baseline and expressed as mean ± SEM; individual values are also represented. Wilcoxon signed-rank test (#) was used to assess statistically the extent of depression. Kruskal–Wallis [H(2,39) = 8.2, p = 0.02] + Dunn’s post-test (*) was used to evaluate significant differences between conditions. ns, non-significant. .

Article Snippet: KBSRC4 , Tocris , #4660.

Techniques: Western Blot, Control, Activation Assay, Activity Assay

Journal: The EMBO Journal

Article Title: Different Ras isoforms regulate synaptic plasticity in opposite directions

doi: 10.1038/s44318-025-00390-8

Figure Lengend Snippet: Reagents and tools table

Article Snippet: KBSRC4 , Tocris , #4660.

Techniques: Recombinant, Plasmid Preparation, Sequencing, Software, Microscopy