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. 2016 Oct 13;17(12):1785–1798. doi: 10.15252/embr.201642416

Figure EV4. Effects of HDAC6 overexpression on the γ‐tubulin at centrosome.

Figure EV4

Related to Figs 5, 6, 7.
  • A, B
    CAMDI overexpression has no effect on tubulin acetylation in the condition of HDAC6 knockdown by shRNA. SH‐SY5Y cells were transfected with indicated vectors, and IP and IB assays were performed using the indicated antibodies. n = 3 independent experiments. N. S., not significant. Two‐way ANOVA followed by Scheffe's post hoc test. Data are presented as mean ± SEM.
  • C
    No significant change in Ac‐HSP90 level in CAMDI‐KO cerebral cortex. Ac‐HSP90 level was assessed by immunoblot analysis. n = 3 independent experiments. One‐way ANOVA followed with Bonferroni's post hoc test. Data are presented as mean ± SEM.
  • D
    HeLa cells were transfected with centrin 2‐EGFP and HDAC6‐HA and immunostained with anti‐EGFP (green) and anti‐γ‐tubulin antibodies (red). DNA was stained with Hoechst 33258. Arrow: HDAC6‐HA and centrin 2‐EGFP transfected cells. Arrowhead: non‐transfected cells. Scale bar, 10 μm.
  • E
    Quantification of the number of Cux1‐positive neurons at P2. n = 3 mice/genotype (WT (vehicle) = 1,426 cells, KO (vehicle) = 2,116 cells, WT (Tubastatin A) = 1,245 cells, KO (Tubastatin A) = 1,311 cells). Two‐way ANOVA followed by Scheffe's post hoc test. Data are presented as mean ± SEM.
  • F
    Three‐chamber social interaction test. The time spent sniffing with stranger mouse. n = 8 for WT (vehicle) mice, n = 6 for KO (vehicle) mice, n = 5 for WT (Tubastatin A) mice, and n = 3 for KO (Tubastatin A) mice. *P < 0.05; two‐way ANOVA followed by Scheffe's post hoc test (main effect of genotype F(1, 18) = 11.11, main effect of drug F(1, 18) = 0.24, interaction F(1, 18) = 0.064). Data are presented as mean ± SEM.