Journal: The Journal of Biological Chemistry

Article Title: Pyrimethamine and a potent analog inhibit NRF2 by suppressing one-carbon metabolism

doi: 10.1016/j.jbc.2025.110659

Figure Lengend Snippet: Pyrimethamine inhibits NRF2 mRNA and protein independently of KEAP1. A . H1299 NQO1-eYFP cells were treated with DMSO vehicle or 10 μM PYR in the presence of PRL295, CDDOme, or sulforaphane. B . Western blot analysis showing dose-dependent effects of PYR on indicated proteins in KYSE70 cells. C . Western blot analysis of PYR (10 μM) time course effects on indicated proteins in KYSE70 cells. Quantitation across biological triplicate experiments is shown below, normalized to VCL (vinculin) (∗ p < 0.05 by one-way ANOVA). D . Western blot analysis of lung and esophageal cancer cells following 48 h treatment with PYR (10 μM). Quantitation across biological triplicate experiments is shown below, normalized to VCL (∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, and ∗∗∗∗ p < 0.0001 by one-way ANOVA). E – F . Western blots assessing the effects of PYR (10 μM; 48 h) on the indicated proteins in KYSE70, KYSE450, or KEAP1 KO derivatives. Quantitation across biological triplicate experiments is shown below (∗∗∗∗ p < 0.0001 by one-way ANOVA). G . Graph showing NRF2 mRNA fold change in KYSE70 cells following 24 and 48 h treatment with PYR (10 μM). NRF2 expression was normalized to RPL13 A. Data are presented as mean ± SD (∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001) by one-way ANOVA (n ≥ 3 biological replicates per group). H . KYSE70 cells harboring a dox-inducible NRF2 shRNA were treated with the indicated amount of dox for 24 h before qPCR and Western blot analysis for NRF2. NRF2 protein was normalized to VCL; NRF2 mRNA expression was normalized to RPL13A. Data are presented as means ± SD (∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001) by one-way ANOVA (n = 3 biological replicates per group). See also .

Article Snippet: KYSE70, KYSE110, KYSE180, and KYSE450 are from DSMZ (Germany), and KYSE450-KEAP1KO and KYSE70-KEAP1KO cells were generated with CRISPR-Cas9.

Techniques: Western Blot, Quantitation Assay, Expressing, shRNA

Journal: Journal of Cellular and Molecular Medicine

Article Title: Multi‐Omics Analysis of Aberrances and Functional Implications of IRF5 in Digestive Tract Tumours

doi: 10.1111/jcmm.70433

Figure Lengend Snippet: Construction of a risk signature using the least absolute shrinkage and selection operator (LASSO) analysis. Partial likelihood deviances for (A) oesophageal squamous cell carcinoma (ESCC) and (B) oesophageal adenocarcinoma (EAC). Coefficient profiles of senescence‐related gene pairs for (C) ESCC and (D) EAC.

Article Snippet: Human ESCC cell lines (KYSE150, KYSE180, KYSE410 and KYSE450) and the human embryonic oesophageal cell line (SHEE) were purchased from iCell Bioscience Inc. (Shanghai, China).

Techniques: Selection

Journal: Journal of Cellular and Molecular Medicine

Article Title: Multi‐Omics Analysis of Aberrances and Functional Implications of IRF5 in Digestive Tract Tumours

doi: 10.1111/jcmm.70433

Figure Lengend Snippet: Kaplan–Meier analysis of high‐ and low‐risk patients (red and blue, respectively) with (A) oesophageal squamous cell carcinoma (ESCC) and (B) oesophageal adenocarcinoma (EAC). Receiver operating characteristic curves for (C) ESCC and (D) EAC. Survival risk curves (top) and sand scatter plots (bottom) for (E) ESCC and (F) EAC.

Article Snippet: Human ESCC cell lines (KYSE150, KYSE180, KYSE410 and KYSE450) and the human embryonic oesophageal cell line (SHEE) were purchased from iCell Bioscience Inc. (Shanghai, China).

Techniques:

Journal: Journal of Cellular and Molecular Medicine

Article Title: Multi‐Omics Analysis of Aberrances and Functional Implications of IRF5 in Digestive Tract Tumours

doi: 10.1111/jcmm.70433

Figure Lengend Snippet: Correlations of immune microenvironments evaluated using ESTIMATE. (A) Immune score and (B) ESTIMATE score for oesophageal squamous cell carcinoma (ESCC). (C) Immune score and (D) ESTIMATE score for oesophageal adenocarcinoma (EAC). Relationships between risk and immune scores for (E) ESCC and (H) EAC. Relationships between risk and ESTIMATE scores for (F) ESCC and (G) EAC.

Article Snippet: Human ESCC cell lines (KYSE150, KYSE180, KYSE410 and KYSE450) and the human embryonic oesophageal cell line (SHEE) were purchased from iCell Bioscience Inc. (Shanghai, China).

Techniques:

Journal: Journal of Cellular and Molecular Medicine

Article Title: Multi‐Omics Analysis of Aberrances and Functional Implications of IRF5 in Digestive Tract Tumours

doi: 10.1111/jcmm.70433

Figure Lengend Snippet: Random forest error rates (left graphs) and relative importance (right graphs) for (A) oesophageal squamous cell carcinoma (ESCC) and (B) oesophageal adenocarcinoma (EAC). Expression of IRF5 and BMI1 in (C) ESCC, (D) EAC and (E) EC.

Article Snippet: Human ESCC cell lines (KYSE150, KYSE180, KYSE410 and KYSE450) and the human embryonic oesophageal cell line (SHEE) were purchased from iCell Bioscience Inc. (Shanghai, China).

Techniques: Expressing

Journal: Journal of Cellular and Molecular Medicine

Article Title: Multi‐Omics Analysis of Aberrances and Functional Implications of IRF5 in Digestive Tract Tumours

doi: 10.1111/jcmm.70433

Figure Lengend Snippet: Expression analyses of IRF5 in four ESCC cell lines using western blotting (A, B). The efficiency of IRF5 ‐knockdown in KYSE150 cells was determined using western blotting (C, D). Ctrl: No siRNA infection; NC: Negative control. Statistical analyses of n = 3 independent experiments were assessed. Results are shown as mean ± SD, ns p ≥ 0.05, * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Human ESCC cell lines (KYSE150, KYSE180, KYSE410 and KYSE450) and the human embryonic oesophageal cell line (SHEE) were purchased from iCell Bioscience Inc. (Shanghai, China).

Techniques: Expressing, Western Blot, Knockdown, Infection, Negative Control