Fig. 1: Suppression of protein synthesis and ribosome stalling in HD cells. | Nature Communications

Fig. 1: Suppression of protein synthesis and ribosome stalling in HD cells.

From: Mutant Huntingtin stalls ribosomes and represses protein synthesis in a cellular model of Huntington disease

Fig. 1

A Representative polysome profile of control, HD-het and HD-homo striatal cells obtained by using sucrose density gradient fractionation. B Quantification of polysome to monosome (PS/MS) ratio in polysome profiles from A (area under the curve). Data are mean ± SEM (control, n = 20; HD-Het, n = 8; HD-Homo, n = 20 independent experiments) ****P < 0.0001 by one-way ANOVA followed by Tukey’s multiple comparison test. C Representative immunoblots of metabolic labeling of protein synthesis, using puromycin, and its quantification (D) in mouse striatal cells. Ponceau staining of the blots was used to quantify the total protein signal in each lane. Data are mean ± SEM (n = 7, independent experiments) ##P < 0.01 by two-tailed Student’s t test and ****P < 0.0001 by one-way ANOVA followed by Tukey’s multiple comparison test. E Representative polysome profiles obtained from control and HD-homo striatal cells at basal (0 min) and after ribosome run-off assay with harringtonine (2 µg/ml, 2, 5, and 8 min). Area e1–e4 shows ribosome movement between control (green) and HD-homo (red) cells. F Quantification of polysome to monosome (PS/MS) ratio in polysome profiles from E (area under the curve). Data are mean ± SEM (at 0 min, n = 8; 2 min, n = 8; 3 min, n = 4; 8 min, n = 6 independent experiments), ***P < 0.001 and ****P < 0.0001, by two-way repeated measures ANOVA, Bonferroni’s multiple comparisons test. G Representative immunoblots of metabolic labeling of protein synthesis, using puromycin, in control and HD-homo striatal cells at basal (0 min) and 2, 5, and 8 min after harringtonine treatment and quantification (H). Ponceau staining of the blots was used to quantify the total protein signal in each lane. Data are mean ± SEM (n = 7, independent experiments), *P < 0.05 and ****P < 0.0001 by two-way repeated measures ANOVA, Bonferroni’s multiple comparisons test. Exact P values are reported in the Source Data file. Source data are provided as a Source Data file.

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