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Fig. 6 | Acta Neuropathologica Communications

Fig. 6

From: Mitochondrial defects in the respiratory complex I contribute to impaired translational initiation via ROS and energy homeostasis in SMA motor neurons

Fig. 6

ROS regulates initiation of protein translation without altering elongation speed. a Scheme of SunRiSE assay. Translation initiation of 10DIV WT or SMA MNs was blocked with 2 µg/ml harringtonine at different time points before adding 10 μg/ml puromycin treatment for 10 min. b, c Neither NAC nor pyruvate changes elongation speed in (b) WT or (c) SMA MNs. Each dot and line represent the average of four independent biological replicates (N = 4). Regression analysis comparing the least square means does not show any significant difference between treatments. d Western blot analysis of p-4EBP1 and 4EBP1 levels in WT (blue circles) and SMA MNs (red squares) (N = 5). ATCB was used as loading control. Each dot represents the data from biological replicates. Two-tailed unpaired t-test was used to determine statistical significance *p < 0.05, **p < 0.01, ***p < 0.001. e, f Western blot analysis of p-4EBP1 and 4EBP1 levels after pharmacological modifications of ROS levels in (e) WT MNs (N = 6) and (f) SMA MNs (N = 6). Each dot represents the data from biological replicates. One-way ANOVA with Tukey HSD post hoc analysis was used to determine statistical significance for multiple comparisons. Bar graphs depict the mean ± S.D. n.s. p > 0.05, *p < 0.05, **p < 0.01, ***p < 0.001

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