sult1b1  (Proteintech)

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: Ischemia onset upregulates H3Y99sulf via the SLC26A1-PAPSS1-SULT1B1 axis (A) Sulfate regulates the H3Y99sulf level in cells. Representative immunoblotting analysis images of triplicated experiments are shown (B and C) OGD treatment upregulates the protein levels of SLC26A1, PAPSS1, and SULT1B1 in cells. Representative immunoblotting analysis images of immortalized cells (SY5Y and HMC3) (B) and primary cells (neurons and microglia) (C) are shown (left panel). The quantitative analyses (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; ∗∗ p < 0.01). (D) The intracellular sulfate concentration is regulated by the OGD treatment. The intracellular sulfate concentration in each sample was normalized to the cell number. Data are presented as the means ± SD ( n = 3). Statistical analyses were performed by two-way ANOVA with Tukey’s multiple comparisons test (∗ p < 0.05, ∗∗ p < 0.01). (E) Depletion of SLC26A1 reduces the H3Y99sulf level in cells. Representative immunoblotting analysis images are shown (left panel). Quantitative analysis results (signals normalized to histone H3) are presented as the means ± SD ( n = 3) (right panel). Statistical analyses were performed by two-tailed Student’s t tests (∗ p < 0.05, ∗∗ p < 0.01). (F) Overexpression of SLC26A1 upregulates the H3Y99sulf level in cells. Representative immunoblotting analysis images are shown (left panel). Quantitative analysis results (signals normalized to histone H3) are presented as the means ± SD ( n = 3) (right panel). Statistical analyses were performed by two-tailed Student’s t tests (∗ p < 0.05). (G) Overexpression of PAPSS1 upregulates the H3Y99sulf level in cells. Representative immunoblotting analysis images of triplicated experiments are shown. (H) Depletion of PAPSS1 downregulates the H3Y99sulf level in cells. Representative immunoblotting analysis images of triplicated experiments are shown. (I) Depletion of PAPSS1 antagonizes sulfate-upregulated H3Y99sulf level in cells. Representative immunoblotting analysis images of triplicated experiments are shown. (J) Depletion of SULT1B1 antagonizes sulfate-upregulated H3Y99sulf level in cells. Representative immunoblotting analysis images of triplicated experiments are shown. (K) The scheme of the SLC26A1-PAPSS1-SULT1B1 mechanism axis regulating the H3Y99sulf level in cells under OGD. The OGD-induced upregulation of SLC26A1 , PAPSS1 , and SULT1B1 are indicated. (L–N) Disruption of the SLC26A1-PAPSS1-SULT1B1 axis significantly diminishes OGD-upregulated H3Y99sulf in cells. Representative immunoblotting analysis images of cells with SLC26A1 (L), PAPSS1 (M), and SULT1B1 (N) depletion are shown (left panel). Quantitative analysis results (signals normalized to histone H3) are presented as the means ± SD ( n = 3) (right panel). Statistical analyses were performed by two-way ANOVA with Tukey’s multiple comparisons test (∗ p < 0.05, ∗∗ p < 0.01). (O) The regulation of SLC26A1-PAPSS1-SULT1B1 in the mouse model of tMCAO. Representative immunoblotting analysis images are shown in the left panel. Quantitative analysis results (H3Y99sulf normalized to histone H3 and other proteins normalized to tubulin) are presented in the right panel ( n = 6). Statistical analyses were performed by two-tailed paired Student’s t tests (∗ p < 0.05, ∗∗ p < 0.01). n indicates biologically independent samples.

Article Snippet: SULT1B1 (human)-shRNA: AACTACAGTGATGGA TCATAG , Sangon Biotech , N/A.

Techniques: Western Blot, Two Tailed Test, Concentration Assay, Over Expression, Disruption

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: SULT1B1 (human)-shRNA: AACTACAGTGATGGA TCATAG , Sangon Biotech , N/A.

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

Journal: Frontiers in Immunology

Article Title: Multi−cohort validation based on coagulation-related genes for predicting prognosis of esophageal squamous cell carcinoma

doi: 10.3389/fimmu.2025.1662599

Figure Lengend Snippet: Identification and construction of the CRGs signature. (A) 19 prognostic CRGs were identified through univariate cox analysis ( p < 0.05). (B) The consensus score matrix of the GSE53625 cohort when k = 2. (C) The CDF curves of consensus matrix for each k, where k ranges from 2 to 6. (D) A heatmap depicted the expression levels of 19 prognostic CRGs, accompanied by clinical characteristic annotations for each cluster. (E) The Kaplan-Meier survival curve depicted significant different overall survival between the two clusters ( p = 0.018). (F) A volcano plot depicted DEGs between the two clusters with criteria of |logFC| > 0.585 and p value < 0.05. (G) Univariate cox regression analysis was conducted to identify prognostic DEGs with a significance level of p < 0.05. (H, I) The coefficient profile of prognostic DEGs was determined by Lasso regression analysis. The optimal λ was achieved when the partial likelihood deviance reached the minimum value. (J) The coefficients of the 6 prognostic DEGs (PTX3, CILP, CFHR4, SULT1B1, IL5RA and FAM151A), which were utilized to construct the CRGs signature, were obtained from multivariate cox analysis. CRGs, coagulation-related genes; DEGs, different expression genes.

Article Snippet: Next, the membrane was incubated with primary antibodies, namely SULT1B1 (1:500; Proteintech, 16050-1-AP, China), E-cadherin (1:5000; Proteintech, 20874-1-AP, China), Vimentin (1:1000; Beyotime, AF1975, China), and GAPDH (1:1000; Servicebio, GB15004-100, China).

Techniques: Expressing, Construct, Coagulation

Journal: Frontiers in Immunology

Article Title: Multi−cohort validation based on coagulation-related genes for predicting prognosis of esophageal squamous cell carcinoma

doi: 10.3389/fimmu.2025.1662599

Figure Lengend Snippet: Low expression of SULT1B1 is associated with poor prognosis in ESCC. (A) In the GSE53625 cohort (tumor samples = 179 and normal samples = 179), the six model CRGs were analyzed with the ‘pROC’ R package. (B, D) The mRNA expression level of SULT1B1 in esophageal cancerous tissues and normal tissues were assessed using the GEPIA and TNMplot databases. (C) The Kaplan-Meier survival curve depicted different overall survival ( p = 0.00021) between the high- and low-SULT1B1 groups using Kaplan-Meier plotter database. (E–I) Boxplots of the difference in the mRNA expression level of SULT1B1 between tumor and normal groups across the GSE20347 , GSE38129 , GSE53625 , GSE53624 , and GSE53622 cohorts. (J–M) The Kaplan-Meier survival curve depicted different overall survival between the high- and low-SULT1B1 groups across the TCGA-ESCC, GSE53625 , GSE53624 , and GSE53622 cohorts. ESCC, esophageal squamous cell carcinoma. *: p < 0.05, **: p < 0.01, ****: p < 0.0001.

Article Snippet: Next, the membrane was incubated with primary antibodies, namely SULT1B1 (1:500; Proteintech, 16050-1-AP, China), E-cadherin (1:5000; Proteintech, 20874-1-AP, China), Vimentin (1:1000; Beyotime, AF1975, China), and GAPDH (1:1000; Servicebio, GB15004-100, China).

Techniques: Expressing

Journal: Frontiers in Immunology

Article Title: Multi−cohort validation based on coagulation-related genes for predicting prognosis of esophageal squamous cell carcinoma

doi: 10.3389/fimmu.2025.1662599

Figure Lengend Snippet: Effects of SULT1B1 on cell proliferation and migration in ESCC cell lines. (A) The expression of SULT1B1 protein in ESCC tissues and pericarcinomatous tissues was detected via western blot. (B) The protein expression levels of SULT1B1 in various ESCC cell lines with statistical analysis. (C) Western blot experiment validated the siRNA knockdown effect in KYSE150 and KYSE410 cells with statistical analysis. (D) Western blot experiment validated the SULT1B1 overexpression in KYSE30 and KYSE410 cells with statistical analysis. (E, H, N, Q) The results of CCK-8 assay in ESCC cells. (F, I, O, R) The effect of knockdown and overexpression of SULT1B1 on the cell cycle of ESCC was detected by flow cytometry. (G, J, P, S) The effect of knockdown and overexpression of SULT1B1 on the apoptosis of ESCC was detected by flow cytometry. (K) The results of scratch wound healing assay of KYSE150 and KYSE410 cells treated with siRNA or negative control of SULT1B1. (L) The results of transwell assay carried out in KYSE150 and KYSE410 cells treated with siRNA or negative control of SULT1B1. (M) Expression of E-cad and Vimentin in si-Ctrl group and si-SULT1B1 group in KYSE150 and KYSE410 cells via western blot. (T) The results of scratch wound healing assay of KYSE30 and KYSE410 cells with SULT1B1 overexpression. (U) The results of transwell assay carried out in KYSE30 and KYSE410 cells with SULT1B1 overexpression. (V) Expression of E-cad and Vimentin in vector group and SULT1B1-OE group in KYSE30 and KYSE410 cells via western blot. ESCC, esophageal squamous cell carcinoma. E-cad, E-cadherin. *: p < 0.05, **: p < 0.01, ***: p < 0.001, ****: p < 0.0001.

Article Snippet: Next, the membrane was incubated with primary antibodies, namely SULT1B1 (1:500; Proteintech, 16050-1-AP, China), E-cadherin (1:5000; Proteintech, 20874-1-AP, China), Vimentin (1:1000; Beyotime, AF1975, China), and GAPDH (1:1000; Servicebio, GB15004-100, China).

Techniques: Migration, Expressing, Western Blot, Knockdown, Over Expression, CCK-8 Assay, Flow Cytometry, Wound Healing Assay, Negative Control, Transwell Assay, Plasmid Preparation