proteins  (Proteintech)

Journal: Chinese Medicine

Article Title: Erianin facilitates pyroptosis in endometrial cancer via targeting m6A reader YTHDF1

doi: 10.1186/s13020-025-01313-9

Figure Lengend Snippet: Erianin targets YTHDF1 directly. A Synthesis of erianin-biotin. B Erianin-biotin 1H NMR spectrum. C CCK-8 results for Ishikawa and HEC-1-B cells incubated with erianin-biotin and erianin (50 and 100 nM) for 24 h. D Schematic workflow of protein array screening to identify erianin targets. E Representative images of the human proteomics chip. Blue, red, and yellow: negative control, positive control, and positive spot, respectively. F Molecular docking simulation of the binding between erianin and YTHDF1. G Thermal stability of erianin and YTHDF1 in EC cells detected using CESTA. H Quantification of YTHDF1 expression. I Expression of YTHDF1 in normal endometrium and EC tissues determined from the CPTAC proteomic database. J Immunohistochemistry (IHC) staining of YTHDF1 in normal endometrium and EC tissues. Scale bar = 20 μm. (K-N) EC cells were incubated with erianin (100 nM). K qRT‒PCR analysis of YTHDF1 expression. L Western blots of YTHDF1 expression. M Ishikawa cells were treated with erianin (100 nM) and cycloheximide (CHX) (50 mg/mL). N Ishikawa cells were treated with MG132 or CQ. O Ubiquitination assay of YTHDF1in the lysates from Erianin treated- EC cells in the presence of MG132 (20 μM). The data are shown as the means ± SD; n = 3; ***P < 0.001 compared with the NC (control group). ns, not significant

Article Snippet: The lysates were incubated with YTHDF1 antibody overnight at 4 °C, followed by addition of protein A/G magnetic beads (HY-K0202, MedChemExpress) and incubation for 4 h at 4 °C.

Techniques: CCK-8 Assay, Incubation, Protein Array, Negative Control, Positive Control, Binding Assay, Expressing, Immunohistochemistry, Western Blot, Ubiquitin Proteomics, Control

Journal: Chinese Medicine

Article Title: Erianin facilitates pyroptosis in endometrial cancer via targeting m6A reader YTHDF1

doi: 10.1186/s13020-025-01313-9

Figure Lengend Snippet: Erianin-induced EC pyroptosis depends on YTHDF1. EC cells were transfected with shYTHDF1 and YTHDF1 vectors. A Survival of EC cells incubated with erianin (100 nM) for 24 h, as detected using CCK-8 assays. B LDH release assay. C , D Western blots of YTHDF1, cleaved GSDME and cleaved caspase-3 in EC cells. E Colony formation assay. F Phase-contrast microscopy images of morphological features. Scale bar, 50 μm. G Representative images of γ-H2AX staining with immunofluorescence. Scale bar, 50 μm. The data are presented as the means ± SD; n = 3, ***P < 0.001 compared with the NC (control group)

Article Snippet: The lysates were incubated with YTHDF1 antibody overnight at 4 °C, followed by addition of protein A/G magnetic beads (HY-K0202, MedChemExpress) and incubation for 4 h at 4 °C.

Techniques: Transfection, Incubation, CCK-8 Assay, Lactate Dehydrogenase Assay, Western Blot, Colony Assay, Microscopy, Staining, Immunofluorescence, Control

Journal: Chinese Medicine

Article Title: Erianin facilitates pyroptosis in endometrial cancer via targeting m6A reader YTHDF1

doi: 10.1186/s13020-025-01313-9

Figure Lengend Snippet: YTHDF1 regulates FOXM1 mRNA stability in EC. A Correlation between YTHDF1 mRNA and FOXM1 mRNA using the GEPIA database. B qRT‒PCR and western blotting experiments were used to validate FOXM1 expression in EC cells transfected with shYTHDF1. C qRT‒PCR and western blotting experiments were used to validate FOXM1 expression in EC cells transfected with shYTHDF1. D , E Relative expression of FOXM1 in Ishikawa and HEC-1-B cells with silenced YTHDF1 incubated with actinomycin D (5 μg/mL). F Results of the RIP assays showing YTHDF1-FOXM1 mRNA binding in the HEC-1-B cells incubated with erianin. G Results of the RIP assays showing YTHDF1-FOXM1 mRNA binding in the HEC-1-B cells with silenced YTHDF1. The data are presented as the mean ± SD; n = 3, *P < 0.05, **P < 0.01, and ***P < 0.001 compared with the NC (control group)

Article Snippet: The lysates were incubated with YTHDF1 antibody overnight at 4 °C, followed by addition of protein A/G magnetic beads (HY-K0202, MedChemExpress) and incubation for 4 h at 4 °C.

Techniques: Western Blot, Expressing, Transfection, Incubation, Binding Assay, Control

Journal: Chinese Medicine

Article Title: Erianin facilitates pyroptosis in endometrial cancer via targeting m6A reader YTHDF1

doi: 10.1186/s13020-025-01313-9

Figure Lengend Snippet: Erianin suppresses EC tumour growth in vivo. A Scheme of the animal experiments. B Erianin inhibits subcutaneous xenograft growth. (n = 6). C Tumor growth curves showing the effects of erianin on EC tumor xenograft growth (n = 6). D Tumor weight. E The protein levels of YTHDF1, caspase-3, cleaved-caspase3, and GSDME in tumor tissues. F Immunohistochemistry (IHC) staining of YTHDF1 in xenografts. Scale bar, 50 μm. G Weight evaluation of mice under Erianin treatment. H Representative images of livers, spleens, and kidneys stained with hematoxylin and eosin (H&E) at the end of the study. Scale bar, 100 μm. I Blood sample biochemical analysis: red blood cells (RBC); white blood cells (WBC); urea (UA); creatinine (CR); aspartate aminotransferase (AST), and alanine aminotransferase (ALT). The data are presented as the means ± SD, n = 6, ***P < 0.001 compared to the NC (control group); ns, not significant

Article Snippet: The lysates were incubated with YTHDF1 antibody overnight at 4 °C, followed by addition of protein A/G magnetic beads (HY-K0202, MedChemExpress) and incubation for 4 h at 4 °C.

Techniques: In Vivo, Immunohistochemistry, Staining, Control