Fig 1.
Body weight of Q175;RhesKO female mice from 21 to 54 weeks of age.
Data are presented as mean ± SEM (WT n = 9; RhesKO n = 11; Q175 n = 9; Q175;RhesKO n = 9); Two-way mixed ANODE followed by Tukey’s test, # p < 0.05, Q175 vs. WT; ¤ p < 0.05, Q175;RhesKO vs. WT). There was no significant difference between Q175 and Q175;RhesKO.
Fig 2.
Discriminant vector bar graph of principal component (PC) analysis.
The vectors represent unique or enriched features (combination of kinematic variables) in Q175 compared to WT group. In the vector bar graph, the bar length and direction correspond to the weight of individual parameters in the corresponding PC.
Fig 3.
Overall gait score in Q175 female mice.
The overall gait score based on differences between Q175 and WT was identified. Scores project normalized parameter data of each mouse onto the discriminant vector; two-way mixed ANOVA followed by Tukey’s test (WT n = 9; RhesKO n = 11; Q175 n = 9; Q175;RhesKO n = 9); Two-way mixed ANOVA followed by Tukey’s test, * p < 0.05, ** p ≤ 0.01, *** p < 0.001.
Fig 4.
MRI analysis of Q175/Rhes female mice at 6–12 months of age.
(A) whole brain volume, (B) striatal volume and (C) cortical volume. Data are presented as mean, + SEM (WT n = 9; RhesKO n = 11; Q175 n = 9; Q175;RhesKO n = 9); Two-way mixed ANODE followed by Tukey’s test, * p < 0.05, Q175 vs. WT andQ175;RhesKO vs. RhesKO.
Fig 5.
MR Spectroscopy analysis of Q175/Rhes female mice, (A) at 6 months of age and (B) at 12 months of age, presenting measured striatal metabolite concentrations from: Ala, alanine; CHO, choline; CR+PCR, creatine+phosphocreatine; CR, creatine; GABA, gamma-aminobutyric acid; Glc, glucose; GLN, glutamine; GLU+GLN, glutamate+glutamine; GLU, glutamate; GPC, glycerophosphocholine; GSH, glutathione; INS, myo-inositol; NAA+NAAG, N-acetylaspartate+N-acetylaspartylglutamate; NAA, N-acetylaspartate; PCh, phosphocholine; PCr, phosphocreatine; TAU, taurine;. Data are presented as mean, + SEM (WT n = 9; RhesKO n = 11; Q175 n = 9; Q175;RhesKO n = 9); Two-way mixed ANODE followed by Tukey’s test, * p < 0.05.
Fig 6.
RhesKO did not restore DARPP32 striatal deficit in mixed gender Q175 mice.
Quantitation of western blots of 5 month striatal samples from male and female mice probed for DARPP32. For each sample, protein level was normalized to in-lane housekeeping protein (vinculin) and presented as percent of WT. Data presented as mean ± SEM (WT n = 6 females, 5 males; RhesKO n = 5 females, 6 males; Q175 n = 6 females, 6 males; Q175;RhesKO n = 6 females, 8 males); One way ANOVA with Bonferroni multiple comparison test; *p<0.05, ****p<0.0001).
Fig 7.
RhesKO did not modulate autophagy in female Q175 mice.
TR-FRET for p62 (A) and lipidated LC3II (B) was performed on 12 month female mouse striatum. Data are expressed as mean ± SEM (WT n = 9; RhesKO n = 11; Q175 n = 9); Q175;RhesKO n = 9; One-way ANOVA statistical analysis: * p <0.05, ** p < 0.01, **** p < 0.0001).
Fig 8.
RhesKO did not modulate HTT levels in female Q175.
MSD for soluble, aggregated and total endogenous mouse HTT was performed on 12 month striatum. Data are expressed as mean ± SEM (WT n = 9; RhesKO n = 11; Q175 n = 9; Q175;RhesKO n = 9); One-way ANOVA statistical analysis: * p <0.05, ** p < 0.01, **** p < 0.0001).
Fig 9.
RhesKO did not modulate mTOR signaling in female Q175.
MSD analysis of p-S6S235/236 normalized by S6 in the 12 month striatum. Data are expressed as mean ± SEM (WT n = 9; RhesKO n = 11; Q175 n = 9; Q175;RhesKO n = 9); One-way ANOVA statistical analysis: **** p < 0.0001).
Fig 10.
Rhes is downregulated in the striatum of mixed gender Q175.
Western blot of striatum from 4M and 14M WT and Q175 mice (A). Quantification of RasGRP1, Rheb and Rhes protein. Data are expressed as mean ± SEM (4M: WT n = 6 male, Q175 n = 5 male and 12-14M: WT n = 9 male, n = 2 female, Q175 n = 9 male, n = 1 female *p<0.05, ****p<0.0001, Student’s t-test.