Fig 2: HT1080 cell lines express endogenous levels of inactive KDACs.(A) qPCR was used to measure compare expression levels of KDAC6 (red) and KDAC8 (blue) in mutant cell lines versus wild-type cells. Each bar represents the relative fold-change calculated using the ΔΔCT method, and error bars represent the standard deviation of at least six replicates. P values are shown for cell lines that demonstrate a significant difference in expression levels compared to wild-type. (B) Immunoblots for KDAC6 and KDAC8 in 50 μg of lysate from each cell line. Actin was probed in 5 μg of the same lysate from each cell line as a loading control. (C) Immunoblots for tubulin and acetylated tubulin in 50 μg of lysate from each cell line. The center panel is an over-exposed version of the top panel to allow comparison of weaker signals. KDAC6 mutant cell lines exhibit increased tubulin acetylation (decreased deacetylation). (D) Immunoblots for SMC3 and acetylated SMC3 in 50 μg of lysate from each cell line. The center panel is an over-exposed version of the top panel to allow comparison of weaker signals. HT1080 KDAC8H143A exhibits increased SMC3 acetylation (decreased deacetylation). (E) Immunoblots for histone H3 and acetylated histone H3 in 50 μg of lysate from each cell line. There is no apparent significant increase in acetylation of histone H3 in the mutant cell lines.

Fig 3: Effect of KDAC6 and KDAC8 catalytic inactivation on gene expression is not equivalent to loss of the KDAC.(A) Venn diagram comparing the number of genes with significant changes in expression level for each cell line perturbing KDAC6 compared to wild-type HT1080 cells. (B) Venn diagram comparing the GO terms associated with gene changes for each KDAC6 mutant line compared to wild-type. (C) Venn diagram comparing the number of genes with significant changes in expression level for HT1080 KDAC8H143A vs HT1080 WT and HAP1 KDAC8KO vs HAP1 WT cells. (D) Venn diagram comparing the GO terms associated with gene changes for HT1080 KDAC8H143A vs HT1080 WT and HAP1 KDAC8KO vs HAP1 WT cells. Gene counts and GO terms are represented in the same way as for Fig 3.

Fig 4: Schematic diagram of the proposed model of LINC01435/YY1/HDAC8 in the diabetic wound healing process schematic diagram of the role of keratinocyte-derived LINC01435 in the dysfunction of human skin endothelial cells under hyperglycemic conditions typical of diabetes

Fig 5: Jurkat cells were treated with indicated concentrations of compound Z16, 6 (A) and SAHA, NC-Z16 (B) for 6 h, respectively. (C) HCT116 cells were treated with indicated concentrations of compound Z16, 6 and SAHA for 6 h. (D) Jurkat and HCT116 cells were treated with indicated concentrations of 32 for 6 h. (E) Jurkat cells were treated with indicated concentrations of compound Z16 for 24 h. (F) THP-1 cells were treated with indicated concentrations of Z16 and 32 for 24 h. The levels of Ac-SMC3, Ac-HH3, Ac-tubulin, and HDAC8 were detected using Western blot. GAPDH was used as a loading control. The levels of Ac-SMC3, Ac-HH3 and HDAC8 were quantified using ImageJ and normalized to the DMSO-treated group. Data are shown as mean ± SEM of two independent experiments. Nonlinear fitting was generated by the GraphPad Prism.ns: not significant, *P < 0.05, **P < 0.01, ***P < 0.001, and ****P < 0.0001 vs DMSO-treated group, one-way analysis of variance (ANOVA).