Journal: Cancer research

Article Title: A search for novel cancer/testis antigens in lung cancer identifies VCX/Y genes expanding the repertoire of potential immunotherapeutic targets

doi: 10.1158/0008-5472.CAN-13-3725

Figure Lengend Snippet: A. Immunohistochemical staining of VCX3A in normal lung (alveolar and bronchus) tissues. Scale bars represent 200 μm.

Article Snippet: A TaqMan polymerase chain reaction (PCR) assay was performed with a 7500 Fast Real-Time PCR System, TaqMan PCR master mix, commercially available primers, FAM-labeled probes for the VCX3A gene (Hs04191277_gH), and VIC-labeled probes for 18S (Hs99999901_S1), according to the manufacturer's instructions (Life Technologies).

Techniques: Immunohistochemical staining, Staining

Journal: Cancer research

Article Title: A search for novel cancer/testis antigens in lung cancer identifies VCX/Y genes expanding the repertoire of potential immunotherapeutic targets

doi: 10.1158/0008-5472.CAN-13-3725

Figure Lengend Snippet: VCX3A expression in NSCLC tissue microarray

Article Snippet: A TaqMan polymerase chain reaction (PCR) assay was performed with a 7500 Fast Real-Time PCR System, TaqMan PCR master mix, commercially available primers, FAM-labeled probes for the VCX3A gene (Hs04191277_gH), and VIC-labeled probes for 18S (Hs99999901_S1), according to the manufacturer's instructions (Life Technologies).

Techniques: Expressing, Microarray, Immunostaining, Mutagenesis

Journal: eBioMedicine

Article Title: Interactive effects of Bisphenol F exposure and TLR4 rs4986790 on unexplained miscarriage

doi: 10.1016/j.ebiom.2025.105968

Figure Lengend Snippet: BPF exposure and TLR4-A transcriptype were associated with women unexplained miscarriage . (A). Schematic diagram for unexplained miscarriage (UM) case–control studies. (B). The median levels of BPF (μg/L) in HC (n = 128) and UM (n = 77) urine samples (p < 0.0001, Student's t-test). (C). Distribution of BPF levels (μg/L) in HC and UM urine samples. (D). Percent change (95% CI) of urinary BPF levels in UM and HC groups. (E). Venn diagram of the intersected mRNAs among the differentially expressed genes (DEGs) in UM vs HC villous tissues, the DEGs in 50 μM BPF-exposed trophoblast cells vs unexposed cells, and the genes containing the miscarriage-related SNPs. (F). Western blot analysis of the protein levels of TLR4 in both HC and UM villous tissues and their relative quantification (n = 24, p < 0.0001, Student's t-test). (G). Percent change (95% CI) of the TLR4 protein levels in UM and HC groups. Data were shown as means ± SD (standard deviation). p < 0.05 meant significant differences compared with control group.

Article Snippet: Six-to-eight-week-old wild-type (WT) C57BL/6 mice (Charles River, Beijing, China) and Tlr4 knockout (KO) mice generated by CRISPR/Cas9 technique (Cyagen Biosciences, China) , were raised in a filter-top cages with a 12 h light/dark cycle, autoclaved bedding, food, and water.

Techniques: Control, Western Blot, Quantitative Proteomics, Standard Deviation

Journal: eBioMedicine

Article Title: Interactive effects of Bisphenol F exposure and TLR4 rs4986790 on unexplained miscarriage

doi: 10.1016/j.ebiom.2025.105968

Figure Lengend Snippet: BPF exposure and TLR4 rs4986790 were interactively associated with miscarriage . (A). The scheme for construction of a BPF-exposed mouse model. (B). The increased weight of BPF-exposed pregnant mice (Δweight = the weight on day 14 –the initial weight on day 1 , n = 6). (C–D). Embryo resorption (indicated by red arrows) in BPF-exposed mice (scale bar, 1 cm) (C) and the average miscarriage rates (embryo resorption rates) in BPF-exposed mice (each n = 6, one-way ANOVA) (D). (E–F). Western blot analysis of murine Tlr4 protein levels in various BPF-exposed mouse placental tissues (E) and their relative quantification (F) (n = 6, one-way ANOVA). (G–H). Western blot analysis of Tlr4 protein levels in placental tissues of BPF-exposed mice with knockdown of murine Tlr4 (G) and its relative quantification (H) (n = 6, one-way ANOVA). (I). HE staining images of mouse placenta of 100 mg/kg/d BPF-exposed mice with knockdown of murine Tlr4. Decidua (D), spongiotrophoblast (SP), labyrinth (LB), and oedema (E) were labelled (scale bar, 300 μm). (J–K). Embryo resorption (indicated by red arrows, scale bar, 1 cm) (J) and the average miscarriage rates (K) in 100 mg/kg/d BPF-exposed mice with knockdown of murine Tlr4 (each n = 6, one-way ANOVA). Data were shown as means ± SD (standard deviation). p < 0.05 meant significant differences compared with control.

Article Snippet: Six-to-eight-week-old wild-type (WT) C57BL/6 mice (Charles River, Beijing, China) and Tlr4 knockout (KO) mice generated by CRISPR/Cas9 technique (Cyagen Biosciences, China) , were raised in a filter-top cages with a 12 h light/dark cycle, autoclaved bedding, food, and water.

Techniques: Western Blot, Quantitative Proteomics, Knockdown, Staining, Standard Deviation, Control

Journal: eBioMedicine

Article Title: Interactive effects of Bisphenol F exposure and TLR4 rs4986790 on unexplained miscarriage

doi: 10.1016/j.ebiom.2025.105968

Figure Lengend Snippet: TLR4-A suppressed trophoblast cell migration/invasion by interacting with β2GPI stronger than TLR4-G . (A). GO analysis of the differentially expressed genes (DEGs) in HTR-8/SVneo-A cells vs HTR-8/SVneo cells. (B). GO analysis of the DEGs in HTR-8/SVneo-A cells vs HTR-8/SVneo-G cells. (C). Transwell assay analysis of the migration/invasion of an identical amount of HTR-8/SVneo, HTR-8/SVneo-A, and HTR-8/SVneo-G cells (scale bar, 100 μm) and their quantification (n = 3, one-way ANOVA). (D–E). The protein levels of TLR4, MMP9, and RAC1 in HTR-8/SVneo, HTR-8/SVneo-A, and HTR-8/SVneo-G cells (D) and their quantification (E) (n = 3, one-way ANOVA). (F). Pearson correlation analysis of the protein levels of TLR4 and MMP9 or RAC1 in HC and UM villous tissues (each n = 12). (G). Western blot analysis of Mmp9 and Rac1 protein levels in placental tissues of BPF-exposed mice with knockdown of murine Tlr4 and their relative quantification (each n = 6, one-way ANOVA). (H). IP assay analysis of the protein levels of β2GPI that was pulled down by TLR4 in HTR-8/SVneo-A and HTR-8/SVneo-G cells, with the protein levels of TLR4, β2GPI shown in cell lysate and their relative quantification (each n = 3, Student's t-test). (I). IP assay analysis of the protein levels of TLR4 that was pulled down by β2GPI in HTR-8/SVneo-A and HTR-8/SVneo-G cells, with the protein levels of TLR4, β2GPI shown in cell lysate and their relative quantification (each n = 3, Student's t-test). Data were shown as means ± SD (standard deviation). p < 0.05 meant significant differences compared with control.

Article Snippet: Six-to-eight-week-old wild-type (WT) C57BL/6 mice (Charles River, Beijing, China) and Tlr4 knockout (KO) mice generated by CRISPR/Cas9 technique (Cyagen Biosciences, China) , were raised in a filter-top cages with a 12 h light/dark cycle, autoclaved bedding, food, and water.

Techniques: Migration, Transwell Assay, Western Blot, Knockdown, Quantitative Proteomics, Standard Deviation, Control

Journal: eBioMedicine

Article Title: Interactive effects of Bisphenol F exposure and TLR4 rs4986790 on unexplained miscarriage

doi: 10.1016/j.ebiom.2025.105968

Figure Lengend Snippet: BPF exposure and TLR4 rs4986790 interactively regulated human trophoblast cell migration/invasion . (A). The number of differentially expressed genes (DEGs) in BPF-exposed HTR-8/SVneo-A or BPF-exposed HTR-8/SVneo-G cells vs their corresponding unexposed cells. (B). GO analysis of the DEGs in BPF-exposed HTR-8/SVneo-A vs unexposed cells. (C). GO analysis of the DEGs in BPF-exposed HTR-8/SVneo-G vs unexposed cells. (D–E). Transwell assay analysis of the migration/invasion of HTR-8/SVneo-A and HTR-8/SVneo-G cells with or without BPF exposure (scale bar, 100 μm) (D) and the quantification of the migrated/invaded cells per view (E) (each n = 3, one-way ANOVA). (F). The reduced number of the migrated/invaded HTR-8/SVneo-A cells or HTR-8/SVneo-G cells after BPF exposure (i.e. the number of migrated/invaded unexposed cells–the number of migrated/invaded BPF-exposed cells) (each n = 3, Student's t-test). (G–I). The protein levels of MMP9 and RAC1 in BPF-exposed HTR-8/SVneo-A and BPF-exposed HTR-8/SVneo-G cells (G) and their relative quantification (H–I) (each n = 3, one-way ANOVA). Data were shown as means ± SD (standard deviation). p < 0.05 meant significant differences compared with control.

Article Snippet: Six-to-eight-week-old wild-type (WT) C57BL/6 mice (Charles River, Beijing, China) and Tlr4 knockout (KO) mice generated by CRISPR/Cas9 technique (Cyagen Biosciences, China) , were raised in a filter-top cages with a 12 h light/dark cycle, autoclaved bedding, food, and water.

Techniques: Migration, Transwell Assay, Quantitative Proteomics, Standard Deviation, Control

Journal: eBioMedicine

Article Title: Interactive effects of Bisphenol F exposure and TLR4 rs4986790 on unexplained miscarriage

doi: 10.1016/j.ebiom.2025.105968

Figure Lengend Snippet: The effects of the interactions of BPF-TLR4 SNP on trophoblast cell migration/invasion . (A). The interaction diagram between BPF and TLR4-A and the enlarged interaction regions. (B). The interaction diagram between BPF and TLR4-G and the enlarged interaction regions. (C). RMSD (Root-mean-square analysis) plots of the equilibrium status of the complex of β2GPI with TLR4-A or TLR4-G in the presence of BPF against stimulation time. (D). The total free binding energy of β2GPI with TLR4-A or TLR4-G in the absence or presence of BPF. (E). IP assay using identical but limited TLR4 antibody showed the protein levels of β2GPI that was pulled down by TLR4 in HTR-8/SVneo-A and HTR-8/SVneo-G cells with or without BPF exposure and their relative quantification (each n = 3, one-way ANOVA). (F). IP assay using identical but limited β2GPI antibody showed the protein levels of TLR4 that was pulled down by β2GPI in HTR-8/SVneo-A and HTR-8/SVneo-G cells with or without BPF exposure and their relative quantification (each n = 3, one-way ANOVA).

Article Snippet: Six-to-eight-week-old wild-type (WT) C57BL/6 mice (Charles River, Beijing, China) and Tlr4 knockout (KO) mice generated by CRISPR/Cas9 technique (Cyagen Biosciences, China) , were raised in a filter-top cages with a 12 h light/dark cycle, autoclaved bedding, food, and water.

Techniques: Migration, Binding Assay, Quantitative Proteomics

Journal: eBioMedicine

Article Title: Interactive effects of Bisphenol F exposure and TLR4 rs4986790 on unexplained miscarriage

doi: 10.1016/j.ebiom.2025.105968

Figure Lengend Snippet: The regulatory mechanisms of BPF effects on TLR4 and β2GPI expression in trophoblast cells . (A). The intersection of the differentially expressed genes in 50 μM BPF-exposed trophoblast cells vs unexposed cells and the potential transcription factors of TLR4 analysed by PROMO software. (B). The mRNA levels of FOXP3 and TLR4 in HTR-8/SVneo-A cells with FOXP3 overexpression or knockdown (each n = 3, Student's t-test (overexpression) and one-way ANOVA (knockdown)). (C–D). The protein levels of FOXP3 and TLR4 in HTR-8/SVneo-A cells with FOXP3 overexpression or knockdown (C) and their relative quantification (D) (each n = 3, Student's t-test (overexpression) and one-way ANOVA (knockdown)). (E). The mRNA levels of FOXP3 in BPF-exposed HTR-8/SVneo-A cells (each n = 3, one-way ANOVA). (F). The protein levels of FOXP3 in BPF-exposed HTR-8/SVneo-A cells and its relative quantification (each n = 3, one-way ANOVA). (G). FOXP3 CHIP assay analysis of the levels of TLR4 promoter region enriched by FOXP3 in HTR-8/SVneo-A cells (each n = 3, Student's t-test). (H). FOXP3 CHIP assay analysis of the levels of TLR4 promoter region enriched by FOXP3 in 100 μM BPF-exposed HTR-8/SVneo-A cells (each n = 3, one-way ANOVA). (I–J). The protein levels of β2GPI in membrane and cytoplasm of BPF-exposed HTR-8/SVneo-A cells (I) and their relative quantification (J) (each n = 3, one-way ANOVA). (K–L). The protein levels of β2GPI in membrane (m-β2GPI) in BPF-exposed HTR-8/SVneo-A or BPF-exposed HTR-8/SVneo-G cells (K) and their relative quantification (L) (each n = 3, one-way ANOVA).

Article Snippet: Six-to-eight-week-old wild-type (WT) C57BL/6 mice (Charles River, Beijing, China) and Tlr4 knockout (KO) mice generated by CRISPR/Cas9 technique (Cyagen Biosciences, China) , were raised in a filter-top cages with a 12 h light/dark cycle, autoclaved bedding, food, and water.

Techniques: Expressing, Software, Over Expression, Knockdown, Quantitative Proteomics, Membrane

Journal: eBioMedicine

Article Title: Interactive effects of Bisphenol F exposure and TLR4 rs4986790 on unexplained miscarriage

doi: 10.1016/j.ebiom.2025.105968

Figure Lengend Snippet: The interactions of BPF-TLR4 SNP in UM and HC villous tissues . (A–B). IP assay using identical but limited TLR4 antibody showed the protein levels of β2GPI that was pulled down by TLR4 in HC and UM villous tissues carrying TLR4-A or -G transcriptypes, with the protein levels of TLR4, β2GPI, MMP9, and RAC1 in tissue lysate (Input) (A) and their relative quantification (B) (UM-A: n = 4, HC-A: n = 4, HC-G: n = 2, one-way ANOVA). (C–D). IP assay using identical but limited β2GPI antibody showed the protein levels of TLR4 that was pulled down by β2GPI in HC and UM villous tissues carrying TLR4-A or -G transcriptypes, with the protein levels of TLR4, β2GPI, MMP9, and RAC1 in tissue lysate (Input) (C) and their relative quantification (D) (UM-A: n = 4, HC-A: n = 4, HC-G: n = 2, one-way ANOVA). (E). The protein levels of TLR4 in lysates of UM-A and HC-A villous tissues (each n = 4, Student's t-test). (F). Pearson correlation analysis of the protein levels of MMP9 or RAC1 and β2GPI that was pulled down by TLR4-A in HC-A villous tissues or by TLR4-G in HC-G villous tissues. (G). Pearson correlation analysis of the protein levels of MMP9 or RAC1 and β2GPI that was pulled down by TLR4-A in HC-A and UM-A villous tissues. (H). Pearson correlation analysis of the protein levels of β2GPI that was pulled down by TLR4-A and urinary BPF levels in HC-A and UM-A groups.

Article Snippet: Six-to-eight-week-old wild-type (WT) C57BL/6 mice (Charles River, Beijing, China) and Tlr4 knockout (KO) mice generated by CRISPR/Cas9 technique (Cyagen Biosciences, China) , were raised in a filter-top cages with a 12 h light/dark cycle, autoclaved bedding, food, and water.

Techniques: Quantitative Proteomics

Journal: eBioMedicine

Article Title: Interactive effects of Bisphenol F exposure and TLR4 rs4986790 on unexplained miscarriage

doi: 10.1016/j.ebiom.2025.105968

Figure Lengend Snippet: The interactions of BPF-TLR4 SNP in BPF-exposed mouse placental tissues . (A). IP assay using identical but limited TLR4 antibody showed the protein levels of β2GPI that was pulled down by Tlr4 in placental tissues of BPF-exposed mice and their relative quantification (each n = 6, one-way ANOVA). (B). IP assay using identical but limited β2GPI antibody showed the protein levels of Tlr4 that were pulled down by β2GPI in placental tissues of BPF-exposed mice and their relative quantification (each n = 6, one-way ANOVA). (C). The protein levels of β2GPI in membrane in placental tissues of BPF-exposed mice and their relative quantification (each n = 6, one-way ANOVA). (D). Nonlinear regression analysis of the association between the protein levels of Mmp9 and β2GPI that was pulled down by Tlr4-A in placental tissues of BPF-exposed mice (each n = 6). (E). Nonlinear regression analysis of the association between the protein levels of Rac1 and β2GPI that was pulled down by Tlr4-A in placental tissues of BPF-exposed mice (each n = 6). (F). The proposed mechanism.

Article Snippet: Six-to-eight-week-old wild-type (WT) C57BL/6 mice (Charles River, Beijing, China) and Tlr4 knockout (KO) mice generated by CRISPR/Cas9 technique (Cyagen Biosciences, China) , were raised in a filter-top cages with a 12 h light/dark cycle, autoclaved bedding, food, and water.

Techniques: Quantitative Proteomics, Membrane