Journal: International Journal of Molecular Sciences

Article Title: FOXC1 Regulates Cytokine Signaling, Inflammatory Pathways, and Retinoid Metabolism to Maintain Limbal Epithelial Cell Homeostasis In Vitro

doi: 10.3390/ijms27041873

Figure Lengend Snippet: Cytochrome P450 (CYP1B1), and paired box protein 6 (PAX6) mRNA and protein levels ( n = 6; n = 5) in primary human limbal epithelial cells (pLECs) 72 h after transfection with control siRNA (blue) or with FOXC1 knockdown siRNA (pink), without or with lipopolysaccharide (LPS) or interleukin 1β (IL-1β) treatment ( A – F ). Data are shown as mean ± SD. Data have been tested for normal distribution using Shapiro–Wilk test. Statistical analysis has been performed using two-way ANOVA followed by Tukey test. p -values below 0.05 were considered statistically significant. Each data point represents an individual donor. FOXC1 siRNA knockdown significantly reduced CYP1B1 mRNA expression compared to siRNA controls under both non-inflammatory and inflammatory conditions (LPS or IL-1β treatment) ( p < 0.001). However, CYP1B1 protein levels did not change significantly following FOXC1 knockdown under any condition ( p ≥ 0.268) ( A – C ). Similarly, FOXC1 siRNA knockdown resulted in decreased PAX6 mRNA expression compared to siRNA controls after LPS treatment ( p = 0.023). In contrast, PAX6 protein levels remained unchanged across all conditions, regardless of inflammatory stimulation ( D – F ).

Article Snippet: CYP1B1 , Rabbit, polyclonal , 61 kDa , 18505-1-AP, Proteintech, Rosemont, Illinois (IL), USA , 1:250.

Techniques: Transfection, Control, Knockdown, 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: 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