Fig. 8: CaMKIIα and Vimentin interact with a specific fragment of the lncRNA SLAMR. | Nature Communications

Fig. 8: CaMKIIα and Vimentin interact with a specific fragment of the lncRNA SLAMR.

From: Synaptically-targeted long non-coding RNA SLAMR promotes structural plasticity by increasing translation and CaMKII activity

Fig. 8

A Schematic of approach to identify protein-protected fragments of SLAMR using a pull-down strategy. B Mapping of seq-Reads of the mouse genome using Salmon and IGV software. C Scheme of cloned SLAMR fragments used to design specific probes. D Representative images of CaMKIIα, Vimentin, and Actin from WB with PD samples. E–F. Plots of WB quantification for CaMKIIα (Independent experiments, N = 11-12, vs Antisense ****p = 4.04E-05, *p = 0.0229; vs sense ##p = 0.0032. One-way ANOVA + Tukey’s test; data as mean ± SEM) and Vimentin (Independent experiments, N = 5-6, One-way ANOVA + Tukey’s test; **p = 0.008, #p = 0.002; data as mean ± SEM). G Schematic of MS2-SLAMR truncated constructs. H–J Quantifications of Supplementary Fig. 7A–F Kymographs. H Deletion of the alternative (92-289) fragment decreased the anterograde speed while deletion of Vimentin/CaMKII (898-1130) binding site increased the speed of MS2-SLAMR:MCP granules compared to the full-length MS2-SLAMR (MS2-SLAMR n = 134 tracks/11 neurons, Δ92-289 n = 56 tracks/9 neurons, Δ898-1130 n = 45 tracks/10 neurons; One-way ANOVA + Tukey’s test; data are presented as min/max). I Deletion of the alternative (92-289) fragment decreased the retrograde speed while deletion of the vimentin/CaMKII (898-1130) binding site led to no change compared to the full-length MS2-SLAMR (MS2-SLAMR n = 80 tracks/11 neurons, Δ92-289 n = 49 tracks/9 neurons, Δ898-1130 n = 39 tracks/10 neurons; One-way ANOVA +Tukey’s test; data are presented as min/max). J Deletion of both the control (92-289) and the vimentin/CaMKII (898-1130) binding site reduced the percent mobile MS2-SLAMR:MCP granules compared to the full-length MS2-SLAMR (MS2-SLAMR n = 11 neurons, Δ92-289 n = 9 neurons, Δ898-1130 n = 10 neurons; One-way ANOVA + Tukey’s test; data as mean ± SEM). K, L Quantifications of Supplementary Fig. 7D–F Kymographs. K Distribution of how MS2-SLAMR, MS2-SLAMRΔ92-289, and MS2-SLAMRΔ898-1130 interact with dendritic spines. L Both the deletions of the alternative fragment and vimentin/CaMKII binding sites decrease the percentage of times that MS2-SLAMR turned around at dendritic spines already occupied by MS2-SLAMR.

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