Fig. 6

AIMTOR reports mTORC1 signaling intensities in primary cultures of hippocampal neurons. a Intensities of the 535nm/450nm ratio in AIMTOR transduced neurons recorded over time after DMSO (control) or rapamycin addition at t = 0. Representative images illustrate this ratio at the indicated times. b Mean BRET intensity recorded before and after transient perfusion of KCl 3 mM-containing ACSF (t = 4 to 10), followed by KCl 55 mM-containing ACSF (t = 36 to 42) to depolarize neurons from − 60 to − 30 mV. Representative images of t = 0 (left) and t = 58 min (right). c, d Basal 535nm/450nm ratio intensities (c) and depolarization-induced BRET increase expressed as percentage of the basal intensity (d), recorded in wild type (WT), Fmr1 KO, and Shank3Δ11 neurons. Each point represents the mean ± SEM for 16 to 30 neurons, 5 regions per neuron. One-way ANOVA multiple comparison statistical analysis for paired (a, b) and unpaired (c) data with *p < 0.05 and ***p < 0.001 compared to the control condition. In a, the initial one-way ANOVA multiple comparison analysis compared within each group the BRET intensity values at each time point to their respective basal BRET value (^###p < 0.001). An additional test based on grouped analyses comparing each time point BRET intensity of the rapamycin versus corresponding DMSO control was carried out with a multiple t test where discovery is determined using the two-stage linear step-up procedure developed by Benjamini and colleagues [32, 33]. BRET intensity is different between rapamycin and DMSO conditions as early as 4 min (arrow *p < 0.05) after adding the drug and remains significantly below the DMSO condition (*p < 0.05, ***p < 0.001) until the end of the monitoring excepted for 3 measures (i.e., 10, 14, and 24 min)