Journal: Cell Reports Medicine

Article Title: Neuroprotective response against the onset of ischemic stroke by upregulation of histone H3Y99 sulfation

doi: 10.1016/j.xcrm.2026.102684

Figure Lengend Snippet: H3Y99sulf promotes glycolysis and sustains cell survival in ischemic stroke (A) Representative H3Y99sulf-enriched gene promoter regions. The y axis represents the fold enrichment level, and the black bar represents the peak detected by MACS2 ( p < 0.01). (B) Validation of the H3Y99sulf enrichments in gene promoter regions. The results of real-time PCR assays following ChIP assays are presented as the means ± SD ( n = 4). Statistical analyses were performed by two-tailed Student’s t tests (∗ p < 0.05, ∗∗ p < 0.01). (C) OGD treatment increases the expression of metabolic genes. Representative immunoblotting images were shown (left panel). Quantitative analysis results (signals normalized to tubulin) are presented as the means ± SD ( n = 3) (right panel). Statistical analyses were performed by two-tailed Student’s t tests (∗ p < 0.05). (D) Disruption of the SLC26A1-PAPSS1-SULT1B1 axis counteracts the regulatory effects of OGD treatment on the protein levels of metabolic genes. Representative immunoblotting images were shown (left panel). Quantitative analysis results (signals normalized to tubulin) are presented as the means ± SD ( n = 3) (right panel). Statistical analyses were performed by two-way ANOVA with Tukey’s multiple comparisons test (n.s. not significant, ∗ p < 0.05, ∗∗ p < 0.01). (E and F) Disruption of the SLC26A1-PAPSS1-SULT1B1 axis counteracts OGD-promoted glycolysis. The normalized glycolytic rate, basal glycolysis, compensatory glycolysis, and the glycolytic reserve were quantitatively analyzed by performing Seahorse assays (E). The time course data are presented as the means ± SEM (E, left panel). The other results are presented as the means ± SD (E, middle and right panels). The quantitative analysis results of lactate production by cells are presented as the means ± SD ( n = 3) (F). Statistical analyses were performed by one-way ANOVA (E) or two-way ANOVA (F) followed by Tukey’s multiple comparisons test (n.s. not significant, ∗ p < 0.05, ∗∗ p < 0.01). (G) Disruption of the H3Y99sulf installation mechanism significantly exacerbates the loss of ATP production in cells treated by OGD. The quantitatively results of ATP in cells are presented as the means ± SD ( n = 3). Statistical analyses were performed by two-way ANOVA with Tukey’s multiple comparisons test (n.s. not significant, ∗ p < 0.05, ∗∗ p < 0.01). (H) Sulfate treatment further increases the protein levels of OGD-upregulated genes. Representative immunoblotting images are shown (left panel). Quantitative analysis results (signals normalized to tubulin) are presented as the means ± SD ( n = 3) (right panel). Statistical analyses were performed by two-way ANOVA with Tukey’s multiple comparisons test (n.s. not significant, ∗ p < 0.05, ∗∗ p < 0.01). (I and J) Sulfate treatment enhances OGD-promoted glycolysis. The normalized glycolytic rate, basal glycolysis, compensatory glycolysis, and the glycolytic reserve were quantitatively analyzed by performing Seahorse assays (I). The time course data are presented as the means ± SEM (I, left panel). The other results are presented as the means ± SD (I, middle and right panels). The quantitative analysis results of lactate production by cells are presented as the means ± SD ( n = 3) (J). Statistical analyses were performed by two-tailed Student’s t tests (I) or two-way ANOVA with Tukey’s multiple comparisons test (J) (n.s. not significant, ∗ p < 0.05). (K) Sulfate treatment significantly increased ATP production in cells treated by OGD. The quantitative results of ATP in cells are presented as the means ± SD ( n = 3). Statistical analyses were performed by two-way ANOVA with Tukey’s multiple comparisons test (n.s. not significant, ∗∗ p < 0.01). (L and M) Disruption of the H3Y99sulf installation mechanism antagonizes the effect of sulfate addition on the glycolysis of the OGD-treated cells. The normalized glycolytic rate, basal glycolysis, compensatory glycolysis, and the glycolytic reserve were quantitatively analyzed by performing Seahorse assays (L). The time course data are presented as the means ± SEM (L, left panel). The other results are presented as the means ± SD (L, middle and right panels). The quantitative analysis results of lactate production by cells are presented as the means ± SD ( n = 3) (M). Statistical analyses were performed by one-way ANOVA (L) or two-way ANOVA (M) followed by Tukey’s multiple comparisons test (n.s. not significant, ∗ p < 0.05). (N) Disruption of the H3Y99sulf installation mechanism antagonizes the effect of sulfate addition on the ATP production in the OGD-treated cells. The quantitative results of ATP in cells are presented as the means ± SD ( n = 3). Statistical analyses were performed by two-way ANOVA with Tukey’s multiple comparisons test (n.s. not significant, ∗ p < 0.05, ∗∗ p < 0.01). n indicates biologically independent samples.

Article Snippet: Quantitative real-time PCR Primer sequence, , Sangon Biotech , N/A.

Techniques: Biomarker Discovery, Real-time Polymerase Chain Reaction, Two Tailed Test, Expressing, Western Blot, Disruption