Fig. 1: Stress-resilient neurons actively repress neuronal function and dopamine synthesis. | Nature Communications

Fig. 1: Stress-resilient neurons actively repress neuronal function and dopamine synthesis.

From: Yin Yang 1 and guanine quadruplexes protect dopaminergic neurons from cellular stress via transmissive dormancy

Fig. 1

a SYTOX green fluorescence in LUHMES cells after a 30-minute thapsigargin (TG) pulse, (ΔF/F0). DD = days of differentiation, D post TG = days after TG removal. Brown-Forsythe and Welch one-way ANOVA with Dunnett’s test using AUC. Mean ± SEM. n = 4 replicates. b Binary firing plots of 80 most active LUHMES cells after TG treatment. Total time of acquisition = 5 min (350 frames). n = 5 for control and 5, 8, 11 for cells 4-, 24-, and 48-hours post-TG, respectively. c Representative traces of the most active neurons per individual recording. Fluo-4 AM fluorescence is normalized to the intrinsic minimum fluorescence per neuron (∆F). d Percentage of neurons with spontaneous calcium transients. Ordinary one-way ANOVA with Šidák’s correction. Mean ± SEM. n = 5 for control and 5, 8, 11 for cells 4-, 24-, and 48-hours post-TG, respectively. e Frequency of neurons with spontaneous calcium transients. Recordings were collected at 1.16 Hz. Ordinary one-way ANOVA with Šidák’s correction. Mean ± SEM. From left to right, n = 124, 74, 80, 56, 62, and 13. f TH protein levels in LUHMES cells 24 h after TG or vehicle treatment as determined by western blot. HEK293t cells serve as an anti-TH control in first lane. g, i qPCR quantification of TH and SLC18A2 transcripts in LUHMES cells after TG treatment. Ordinary one-way ANOVA with Šidák’s correction. Mean ± SEM. n = 4 for 3- and 6-hours post-TG and n = 7 for 24-hours post-TG. h Dopamine content in LUHMES cells determined by ELISA after TG treatment. Kruskal-Wallis ANOVA with Dunn’s correction. Mean ± SEM. n = 3 replicates. j, k qPCR quantification of TH and SLC18A2 transcripts in LUHMES cells treated with 100 µM paraquat (PQ) for 3 h. Unpaired two-tailed t-test. Mean ± SEM. n = 3 replicates. l Whole-brain RNA-seq collected 2 h and 7 days post-TBI in mice (GSE173431). Transcripts are normalized to time-matched uninjured controls. Ordinary one-way ANOVA with Šidák’s correction. Mean ± SEM. n = 3 for 2 h control and 7 d injury. n = 4 for 2 h injury and 7 d control.

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