Journal: Oncology reports

Article Title: Targeting CDK9: A novel biomarker in the treatment of endometrial cancer.

doi: 10.3892/or.2020.7746

Figure Lengend Snippet: Figure 3. CDK9 expression in endometrial cancer cell lines. (A) Expression levels of CDK9 in endometrial cancer cell lines (AN3CA, ARK‑2, HEC‑1A, HEC‑1B, lshikawa, RL95‑2 and SPAC1S) as determined by western blotting. (B) Relative expression of CDK9 and α‑tubulin in the endometrial cancer cell lines. CDK9, cyclin‑dependent kinase 9.

Article Snippet: AN3CA was obtained from the German Collection of Microorganisms and Cell Cultures (DSMZ; Braunschweig, Germany).

Techniques: Expressing, Western Blot

Journal: Oncology reports

Article Title: Targeting CDK9: A novel biomarker in the treatment of endometrial cancer.

doi: 10.3892/or.2020.7746

Figure Lengend Snippet: Figure 5. CDK9 inhibitor reduces endometrial cancer cell proliferation by suppressing transcription elongation and inducing apoptosis in endometrial cancer cells. (A and B) Relative cell viability of AN3CA and SPAC1S cells after exposure to different concentrations of the CDK9 inhibitor LDC067 for 5 days. **P<0.01 compared with the lowest concentration group (1x10‑3 µM). (C and D) Expression levels of CDK9 and related signaling pathway proteins involved in transcription and apoptosis after treatment with LDC067 in cells by western blot analysis. CDK9, cyclin‑dependent kinase 9; Mcl‑1, myeloid cell leukemia‑1; Bax, proapoptotic protein BCL2 associated X, apoptosis regulator; PARP, poly(ADP‑ribose) polymerase.

Article Snippet: AN3CA was obtained from the German Collection of Microorganisms and Cell Cultures (DSMZ; Braunschweig, Germany).

Techniques: Concentration Assay, Expressing, Western Blot

Journal: Oncology reports

Article Title: Targeting CDK9: A novel biomarker in the treatment of endometrial cancer.

doi: 10.3892/or.2020.7746

Figure Lengend Snippet: Figure 4. CDK9 knockdown by siRNA transfection suppresses endometrial cancer cell proliferation. (A and B) MTT assay revealed significant dose‑dependent inhibition of cell proliferation after CDK9 siRNA treatment. **P<0.01 compared with the cell only control group. (C and D) Expression levels of CDK9 and related signaling pathway proteins involved in transcription and apoptosis after transfection of CDK9 siRNA and nonspecific siRNA (NC siRNA) in AN3CA and SPAC1S cell lines by western blot analysis. CDK9, cyclin‑dependent kinase 9; Mcl‑1, myeloid cell leukemia‑1; Bax, proapoptotic protein BCL2 associated X, apoptosis regulator.

Article Snippet: AN3CA was obtained from the German Collection of Microorganisms and Cell Cultures (DSMZ; Braunschweig, Germany).

Techniques: Knockdown, Transfection, MTT Assay, Inhibition, Control, Expressing, Western Blot

Journal: Oncology reports

Article Title: Targeting CDK9: A novel biomarker in the treatment of endometrial cancer.

doi: 10.3892/or.2020.7746

Figure Lengend Snippet: Figure 7. Inhibition of CDK9 reduces endometrial cancer cell migration. (A and B) Representative images of AN3CA and SPAC1S cell migration after CDK9 inhibitor LDC067 treatment for 0, 24, and 48 h. (C and D) Quantification of cell migration distance of AN3CA and SPAC1S cells after LDC067 treatment. **P<0.01 compared with the Cell only group. CDK9, cyclin‑dependent kinase 9.

Article Snippet: AN3CA was obtained from the German Collection of Microorganisms and Cell Cultures (DSMZ; Braunschweig, Germany).

Techniques: Inhibition, Migration

Journal: Oncology reports

Article Title: Targeting CDK9: A novel biomarker in the treatment of endometrial cancer.

doi: 10.3892/or.2020.7746

Figure Lengend Snippet: Figure 6. Inhibition of CDK9 suppresses endometrial cancer cell colony formation. (A) Representative images of endometrial cancer cell colony formation after incubation with different concentrations of LDC067 (0, 2.5, 5.0, and 10 µM) for 14 days. (B and C) Quantification of clonogenicity formation of AN3CA (B) and SPAC1S (C) cells after LDC067 treatment. **P<0.01 compared with the Cell only group. CDK9, cyclin‑dependent kinase 9.

Article Snippet: AN3CA was obtained from the German Collection of Microorganisms and Cell Cultures (DSMZ; Braunschweig, Germany).

Techniques: Inhibition, Incubation

Journal: Reproductive Biology and Endocrinology : RB&E

Article Title: Uterine Foxl2 regulates the adherence of the Trophectoderm cells to the endometrial epithelium

doi: 10.1186/s12958-018-0329-y

Figure Lengend Snippet: Foxl2 expression in uterine cell lines. qRT-PCR of Foxl2 mRNA expression in AN3-CA and Ishikawa endometrial cell lines. Endogenous expression in Ishikawa cells is significantly lower

Article Snippet: The AN3-CA cells, non-receptive human endometrial cells (cell line obtained from CLS Cell Line Services GmbH, Eppelheim, Germany), were grown in MEM (Biological Industries, Israel) with 10% fetal bovine serum (FBS, Hyclone, Biological Industries, Israel), Ishikawa cells, receptive human endometrial cells (cell line obtained from ATCC, Manassas, VA), were grown in DMEM (Biological Industries, Israel) with 10% FBS.

Techniques: Expressing, Quantitative RT-PCR

Journal: Reproductive Biology and Endocrinology : RB&E

Article Title: Uterine Foxl2 regulates the adherence of the Trophectoderm cells to the endometrial epithelium

doi: 10.1186/s12958-018-0329-y

Figure Lengend Snippet: Foxl2 effects in an in vitro model of attachment. a Mouse embryo attachment is higher after Foxl2 knockdown in AN3-CA cells. b Overexpression of Foxl2 in Ishikawa cells reduces mouse embryo attachment. c Jeg3 spheroid attachment is higher after Foxl2 knockdown in AN3-CA. d Overexpression of Foxl2 in Ishikawa cells decreases Jeg3 spheroid attachment. * p < 0.05

Article Snippet: The AN3-CA cells, non-receptive human endometrial cells (cell line obtained from CLS Cell Line Services GmbH, Eppelheim, Germany), were grown in MEM (Biological Industries, Israel) with 10% fetal bovine serum (FBS, Hyclone, Biological Industries, Israel), Ishikawa cells, receptive human endometrial cells (cell line obtained from ATCC, Manassas, VA), were grown in DMEM (Biological Industries, Israel) with 10% FBS.

Techniques: In Vitro, Knockdown, Over Expression

Journal: Reproductive Biology and Endocrinology : RB&E

Article Title: Uterine Foxl2 regulates the adherence of the Trophectoderm cells to the endometrial epithelium

doi: 10.1186/s12958-018-0329-y

Figure Lengend Snippet: Foxl2 effect on the expression of Wnt/Fzd family members. a , d , g The expression of Fzd6 and Wnt11 is higher, while the expression of Kremen2 , a Wnt/Fzd family inhibitor, is lower in receptive endometrial Ishikawa cells, which express low Foxl2 levels, compared to AN3-CA endometrial non-receptive cells, which express higher levels of Foxl2 . b , e , h Knockdown of Foxl2 in AN3-CA leads to an increase in Fzd6 and Wnt11 expression and a reduction in Kremen2 levels. c , f , i Overexpression of Foxl2 in Ishikawa cells leads to a decrease in Fzd6 and Wnt11 levels but elevates Kremen2 levels. * p < 0.05

Article Snippet: The AN3-CA cells, non-receptive human endometrial cells (cell line obtained from CLS Cell Line Services GmbH, Eppelheim, Germany), were grown in MEM (Biological Industries, Israel) with 10% fetal bovine serum (FBS, Hyclone, Biological Industries, Israel), Ishikawa cells, receptive human endometrial cells (cell line obtained from ATCC, Manassas, VA), were grown in DMEM (Biological Industries, Israel) with 10% FBS.

Techniques: Expressing, Knockdown, Over Expression

Journal: Reproductive Biology and Endocrinology : RB&E

Article Title: Uterine Foxl2 regulates the adherence of the Trophectoderm cells to the endometrial epithelium

doi: 10.1186/s12958-018-0329-y

Figure Lengend Snippet: Foxl2 effect on the expression of genes involved in apoptosis. a , d , g The expression of Tnfip3 and Atf3 is elevated, while the expression of Ier3 is decreased, in receptive endometrial Ishikawa cells, expressing low Foxl2 levels as compared to AN3-CA, endometrial non-receptive cells, expressing high levels of Foxl2 . b , e , h Tnfip3 and Atf3 expression is reduced, while the expression of Ier3 is elevated in AN3-CA after Foxl2 knockdown. c , f , i Overexpression of Foxl2 in Ishikawa cells is associated with a decrease in Tnfip3 and Atf3 levels and increase in Ier3 levels. * p < 0.05

Article Snippet: The AN3-CA cells, non-receptive human endometrial cells (cell line obtained from CLS Cell Line Services GmbH, Eppelheim, Germany), were grown in MEM (Biological Industries, Israel) with 10% fetal bovine serum (FBS, Hyclone, Biological Industries, Israel), Ishikawa cells, receptive human endometrial cells (cell line obtained from ATCC, Manassas, VA), were grown in DMEM (Biological Industries, Israel) with 10% FBS.

Techniques: Expressing, Knockdown, Over Expression

Journal: Reproductive Biology and Endocrinology : RB&E

Article Title: Uterine Foxl2 regulates the adherence of the Trophectoderm cells to the endometrial epithelium

doi: 10.1186/s12958-018-0329-y

Figure Lengend Snippet: Foxl2 effect on the expression of genes involved in embryo implantation. The expression of Rgs2 and Cxcl1 is elevated in receptive endometrial Ishikawa cells, expressing low Foxl2 levels as compared to AN3-CA, endometrial non-receptive cells, expressing high levels of Foxl2 , ( a , d ) as well as in AN3-CA Foxl2 depleted cells ( b , e ). Overexpression of Foxl2 in Ishikawa cells resulted in a decrease in Rgs2 and Cxcl1 levels ( c , f ). * p < 0.05

Article Snippet: The AN3-CA cells, non-receptive human endometrial cells (cell line obtained from CLS Cell Line Services GmbH, Eppelheim, Germany), were grown in MEM (Biological Industries, Israel) with 10% fetal bovine serum (FBS, Hyclone, Biological Industries, Israel), Ishikawa cells, receptive human endometrial cells (cell line obtained from ATCC, Manassas, VA), were grown in DMEM (Biological Industries, Israel) with 10% FBS.

Techniques: Expressing, Over Expression

Journal: Oncotarget

Article Title: SOX17 is a tumor suppressor in endometrial cancer

doi: 10.18632/oncotarget.12582

Figure Lengend Snippet: Real-time PCR and western blot assays were used to characterize MAML3 expression in SOX17-silenced AN3CA A. HEC-1B B. and Ishikawa C. cells and in SOX17-overexpressing AN3CA D. HEC-1B E. and Ishikawa F. cells. SOX17 putative binding site located −803 to −809 bp from the transcription start site in the MAML3 promoter region. G. ChIP analysis of SOX17 binding to MAML3 in HEC-1B cells H.

Article Snippet: The human EC cell lines Ishikawa, HEC-1B and AN3CA, were obtained from and maintained as recommended by the China Center for Type Culture Collection (CCTCC).

Techniques: Real-time Polymerase Chain Reaction, Western Blot, Expressing, Binding Assay

Journal: Oncology Research

Article Title: Regulation of Histone Emulsification by HPDL via LDHA/LDHB Promotes EC Cell Proliferation

doi: 10.32604/or.2026.068833

Figure Lengend Snippet: Knockdown of HPDL decreased the level of histone lactonization modification in EC cells. ( A ) Western blot (WB) analysis confirming the expression of histone lactate dehydrogenase (HLD) antibodies in AN3CA cells with HPDL knockdown; ( B ) WB analysis confirming the expression of HLD antibodies in HEC-1-B cells with HPDL knockdown; ( C ) Quantitative polymerase chain reaction (qPCR) analysis of HPDL expression following triple-sequence plasmid knockdown in AN3CA cells; ( D ) qPCR analysis of HPDL expression following triple-sequence plasmid knockdown in HEC-1-B cells; ( E ) Measurement of lactate content in AN3CA cells subsequent to HPDL knockdown; ( F ) Measurement of lactate content in HEC-1-B cells subsequent to HPDL knockdown. ( G ) Green fluorescence signifies HPDL protein expression, while blue fluorescence indicates immunofluorescence staining of cell nuclei; scale bar is 50 μm; ( H ) Expression levels of HPDL, Pan Kla, and H3K18la after the addition of varying concentrations of sodium lactate to AN3CA Si HPDL knockdown cell lines; scale bar represents 50 μm. Expression levels of HPDL, Pan Kla, and H3K18la in AN3CA Si HPDL knockdown cell lines with the addition of varying concentrations of sodium lactate; scale bar represents 50 μm; ( I ) Expression levels of HPDL, Pan Kla, and H3K18la after the addition of varying concentrations of sodium lactate to AN3CA normal cell lines. *indicates p < 0.05, **indicates p < 0.01, ***indicates p < 0.001

Article Snippet: AN3CA cell line , Procell , CL-0530.

Techniques: Knockdown, Modification, Western Blot, Expressing, Real-time Polymerase Chain Reaction, Sequencing, Plasmid Preparation, Fluorescence, Immunofluorescence, Staining

Journal: Oncology Research

Article Title: Regulation of Histone Emulsification by HPDL via LDHA/LDHB Promotes EC Cell Proliferation

doi: 10.32604/or.2026.068833

Figure Lengend Snippet: The level of histone lactylation was elevated in EC cells overexpressing HPDL ( A ) Expression levels of HPDL, Pan Kla, and H3K18la following the overexpression of HPDL in the HEC-1-B cell line; ( B ) Expression levels of HPDL, Pan Kla, and H3K18la after treating the HEC-1-B cell LV-HPDL with varying concentrations of sodium oxalate; ( C ) Overexpression and knockdown strains of AN3CA; ( D ) Lactate levels in HEC-1-B overexpression and knockdown strains; ( E ) Immunofluorescence of AN3CA cells for quantifying H3K18la immunofluorescence across different groups; ( F ) Immunofluorescence of AN3CA cells for quantifying DAPI immunofluorescence across different groups; ( G ) Immunofluorescence of AN3CA cells for quantifying the expression of H3K18la and DAPI, scale bar is 50 μm; *indicating p < 0.05, **indicating p < 0.01, ***indicating p < 0.001, and ns indicating no statistical significance

Article Snippet: AN3CA cell line , Procell , CL-0530.

Techniques: Expressing, Over Expression, Knockdown, Immunofluorescence

Journal: Oncology Research

Article Title: Regulation of Histone Emulsification by HPDL via LDHA/LDHB Promotes EC Cell Proliferation

doi: 10.32604/or.2026.068833

Figure Lengend Snippet: The expression of HPDL promotes the proliferation of EC cells and affects the cell cycle. ( A ) CCK-8 profiles in AN3CA cells; ( B ) CCK-8 profiles in HEC-1-B cells; ( C ) Quantitative results of plate cloning assay of HEC-1-B cells; ( D ) Plate cloning assay of AN3CA and HEC-1-B cells; ( E ) Quantification results of plate cloning of AN3CA cells; ( F ) AN3CA cell cycle assay, *indicating p < 0.05 and ***indicating p < 0.001

Article Snippet: AN3CA cell line , Procell , CL-0530.

Techniques: Expressing, CCK-8 Assay, Cloning, Cell Cycle Assay

Journal: Oncology Research

Article Title: Regulation of Histone Emulsification by HPDL via LDHA/LDHB Promotes EC Cell Proliferation

doi: 10.32604/or.2026.068833

Figure Lengend Snippet: HPDL promotes the migration and invasion of EC cells. ( A ) Cell scratch assay of the normal, overexpression and knockdown groups of the HPDL gene in AN3CA cells, scale bar is 50 μm; ( B ) Cell scratch assay of the normal, overexpression and knockdown groups of the HPDL gene in HEC-1-B cells; ( C ) AN3CA and HEC-1-B cells with the normal, overexpression and knockdown groups of the HPDL gene in the quantitative histograms of cell scratch assay cells; ( D ) quantitative histograms of Transwell migration assay of AN3CA cells; ( E ) quantitative histograms of Transwell migration assay of HEC-1-B cells; ( F ) Transwell migration assay of AN3CA and HEC-1-B cells, scale bar is 50 μm; ( G ) AN3CA and HEC-1-B cells Transwell invasion assay; ( H ) AN3CA and HEC-1-B cell HPDL overexpression Transwell invasion assay; ( I ) Transwell invasion assay with HPDL knockdown in AN3CA and HEC-1-B cells, ***indicating p < 0.001

Article Snippet: AN3CA cell line , Procell , CL-0530.

Techniques: Migration, Wound Healing Assay, Over Expression, Knockdown, Transwell Migration Assay, Transwell Invasion Assay

Journal: Oncology Research

Article Title: Regulation of Histone Emulsification by HPDL via LDHA/LDHB Promotes EC Cell Proliferation

doi: 10.32604/or.2026.068833

Figure Lengend Snippet: HPDL promotes histone lactylation levels in EC cells by upregulating LDHA/LDHB expression. ( A ) Chordal graph of GO/KEGG analysis; ( B ) Western blot (WB) demonstrates detection of Pan Kla, H3K18la antibody expression after LDHA and LDHB inhibition; ( C ) Reverse transcription-quantitative polymerase chain reaction (qPCR) showing the expression of HPDL in AN3CA cells with SiNC, SiLDHA, and SiLDHB; ( D ) WB demonstration of fluorescence transfection efficiency and LDHA/LDHB and HPDL expression in AN3CA cells with SiNC, SiLDHA, and SiLDHB; ( E ) qPCR of AN3CA cells with decreased LDHA/LDHB expression in the SiHPDL group, and decreased LDHA/LDHB expression in the LVHPDL group, where LDHA/LDHB expression increased; ( F) H3K18la expression in the LDHA promoter region in AN3CA cells with SiNC, LV-HPDL, and LV-HPDL+Oxmate; ( G ) H3K18la expression in the LDHB promoter region in AN3CA cells with SiNC, LV-HPDL, and LV-HPDL+Oxmate; ( H ) H3K18la expression in the LDHA promoter region in AN3CA cells with SiNC, SiHPDL, and SiHPDL+L-NaLa; ( I ) H3K18la expression in the LDHB promoter region in AN3CA cells with SiNC, SiHPDL, and SiHPDL+L-NaLa; *indicates p < 0.05, **indicates p < 0.01, ***indicates p < 0.001

Article Snippet: AN3CA cell line , Procell , CL-0530.

Techniques: Expressing, Western Blot, Inhibition, Reverse Transcription, Real-time Polymerase Chain Reaction, Fluorescence, Transfection

Journal: Biology Direct

Article Title: Platelet-activating factor induces ferroptosis by binding to ATF3 and inhibiting the SLC7A11/GPX4 axis to suppress the progression of endometrial carcinoma

doi: 10.1186/s13062-025-00713-z

Figure Lengend Snippet: PAF inhibits EC cell proliferation in AN3CA and HEC1B cells in vitro . (A , B) The IC 50 values of PAF in AN3CA ( A ) and HEC1B ( B ) cells were determined using cell viability assays. One-way ANOVA with Tukey’s post hoc test was applied. (C , D) The colony-forming potential of AN3CA and HEC1B cells was assessed by crystal violet staining after incubation with 60 µM PAF. (E , F) PAF (60 µM) was incubated with AN3CA ( E ) and HEC1B ( F ) cells in a glass-bottom cell culture dish for 48 h. Proliferating cells were examined via EdU incorporation assay (orange) and Hoechst 33,342 (blue) nuclear staining for EC cells, and the images were merged. Scale bars, 100 μm. (G , H) Patient-derived EC organoids were treated with different concentrations of PAF (0, 20, 40, and 100 µM) for 48 h. Cell viability was measured using an ATP assay kit. Representative images ( G ) and statistical data ( H ) are shown. Scale bars, 200 μm. One-way ANOVA with Dunnett’s multiple comparisons test was used. (I) Representative transmission electron microscopy images show the mitochondrial morphology in AN3CA and HEC1B cells treated with control and PAF (60 µM) for 48 h. Scale bars: 2 μm (Black) or 1 μm (Green). ∗, p < 0.05; ∗∗∗, p <0.001

Article Snippet: HEC-1B (STCC10605P), AN3CA (STCC10601P), and 293T (STCC10301P) cell lines were purchased from Servicebio (Hubei, China) and were recently authenticated by Short Tandem Repeat profiling.

Techniques: In Vitro, Staining, Incubation, Cell Culture, Derivative Assay, ATP Assay, Transmission Assay, Electron Microscopy, Control

Journal: Biology Direct

Article Title: Platelet-activating factor induces ferroptosis by binding to ATF3 and inhibiting the SLC7A11/GPX4 axis to suppress the progression of endometrial carcinoma

doi: 10.1186/s13062-025-00713-z

Figure Lengend Snippet: PAF induces ferroptosis in EC cells. (A , B) Gene set enrichment analysis plots of the RNA-Seq analysis of ferroptosis-related gene modules in AN3CA (A) and HEC1B (B) cells treated with PAF. NES, normalized enrichment score. (C , D) Volcano plots of differentially expressed genes in the custom gene set. (E , F) Flow cytometry analysis of ROS levels in AN3CA (E) and HEC1B (F) cells treated with PAF for 48 h. (G) Intracellular Fe 2+ intensity assays in AN3CA and HEC1B cells treated with PAF for 48 h. (H) BODIPY-C11 staining of AN3CA and HEC1B cells treated with PAF (60 µM) for 8 h. Scale bars, 100 μm. (I , J) Lipid peroxidation was measured using the BODIPY 581/591 C11 fluorescent probe. (K) WB analysis of the expression of SLC7A11, GPX4, and FTH1 in AN3CA and HEC1B cells treated with different concentrations (20, 40, and 60 µM) of PAF for 48 h. (L , M) AN3CA (L) and HEC1B (M) cell viability treated with a combination of PAF (60 µM) and the ferroptosis inhibitor ferrostatin-1 (Ferr-1, 10 µM). * p < 0.05, ** p < 0.01 and *** p < 0.001

Article Snippet: HEC-1B (STCC10605P), AN3CA (STCC10601P), and 293T (STCC10301P) cell lines were purchased from Servicebio (Hubei, China) and were recently authenticated by Short Tandem Repeat profiling.

Techniques: RNA Sequencing, Flow Cytometry, Staining, Expressing

Journal: Biology Direct

Article Title: Platelet-activating factor induces ferroptosis by binding to ATF3 and inhibiting the SLC7A11/GPX4 axis to suppress the progression of endometrial carcinoma

doi: 10.1186/s13062-025-00713-z

Figure Lengend Snippet: PAF induces ferroptosis via the ATF3/GPX4 and ATF3/SLC7A11 axes. (A) Heat map representing the significantly regulated transcription factors detected via RNA-seq analysis of AN3CA and HEC1B cells. (B , C) The mRNA expression levels of ATF3 and DDIT3 were in AN3CA and HEC1B cells treated with PAF (60 µM) for 48 h. (D , E) Protein expression of ATF3 in AN3CA and HEC1B cells treated with PAF (0, 20, 40, and 60 µM) for 48 h (D), and the quantifiable data are shown (E). (F) Confocal microscopy of AN3CA and HEC1B cells in the presence of PAF (60 µM). Cells were stained for ATF3 (green) and phalloidin (red). DAPI was used to visualize the nucleus (blue). Scale bar, 50 μm (left) or 100 μm (right). (G) The potential binding sites between ATF3 and GPX4/SLC7A11 were predicted by JASPAR. (H) Schematic diagram of ATF3 binding site-mutated SLC7A11 reporter vector (pro-SLC7A11-mut) and GPX4 reporter vector (pro-GPX4-mut). (I , J) Dual luciferase assay of the luciferase activity of AN3CA (I) and HEC1B (J) cells. (K) Immunoblotting analysis of the expression of GPX4 and SLC7A11 following ATF3 overexpression in AN3CA and HEC1B cells. (L) Immunoblotting analysis of the protein expression of SLC7A11, GPX4, and ATF3 after ATF3 silencing with siRNA coupled with PAF treatment in AN3CA cells. * p < 0.05, ** p < 0.01 and *** p < 0.001, ns . not significant

Article Snippet: HEC-1B (STCC10605P), AN3CA (STCC10601P), and 293T (STCC10301P) cell lines were purchased from Servicebio (Hubei, China) and were recently authenticated by Short Tandem Repeat profiling.

Techniques: RNA Sequencing, Expressing, Confocal Microscopy, Staining, Binding Assay, Plasmid Preparation, Luciferase, Activity Assay, Western Blot, Over Expression

Journal: Biology Direct

Article Title: Platelet-activating factor induces ferroptosis by binding to ATF3 and inhibiting the SLC7A11/GPX4 axis to suppress the progression of endometrial carcinoma

doi: 10.1186/s13062-025-00713-z

Figure Lengend Snippet: PAF binds to ATF3 and reduces its ubiquitination. (A , B) Molecular docking indicates the binding details between PAF and ATF3. The surface representation of the protein residues ( A ) and 2D representation of the binding interaction of PAF and ATF3 ( B ) are depicted. (C , D) Surface plasmon resonance of the affinity of anti-ATF3 antibody ( C ) and PAF ( D ) for ATF3 protein. K D , dissociation constant. (E) WB analysis shows that PAF stabilized ATF3 across different temperature gradients in the CETSA in 293T cells. (F) WB analysis indicates that PAF promoted the resistance of ATF3 to pronase digestion in the DARTS assay in 293T cells. (G) CHX chase analysis of ATF3 protein expression after treatment with PAF in AN3CA and HEC1B cells. (H) WB analysis of ATF3 in ATF3 -overexpressing AN3CA and HEC1B cells. The cells were pretreated with PAF (60 µM) for 24 h and then treated with CHX and MG132 for 24 h. (I) Representative WB images demonstrate the ubiquitination of ATF3 in 293T cells co-transfected with ATF3-Flag, HA-Ub, and plasmids for 24 h. Cellular lysates were collected after 3 h of treatment with PAF, purified with a Flag-tag protein purification kit, and then subjected to WB with anti-HA and anti-ATF3

Article Snippet: HEC-1B (STCC10605P), AN3CA (STCC10601P), and 293T (STCC10301P) cell lines were purchased from Servicebio (Hubei, China) and were recently authenticated by Short Tandem Repeat profiling.

Techniques: Ubiquitin Proteomics, Binding Assay, SPR Assay, Expressing, Transfection, Purification, FLAG-tag, Protein Purification

Journal: Biology Direct

Article Title: Platelet-activating factor induces ferroptosis by binding to ATF3 and inhibiting the SLC7A11/GPX4 axis to suppress the progression of endometrial carcinoma

doi: 10.1186/s13062-025-00713-z

Figure Lengend Snippet: Treatment with PAF inhibits EC progression by promoting ferroptosis in vivo. (A) Timeline of the nude mice experiments. Mice bearing AN3CA and HEC1B subcutaneous tumors were injected with PAF every 3 days for a total of 6 times. (B) The resected tumors. (C , D) Volume ( C ) and weight ( D ) of tumors. The negative control group received a saline solution (vehicle). Data are presented as means ± SEM and were analyzed using the Student’s t -test was applied ( n = 5 in the AN3CA group; n = 6 in the HEC1B group). (E) Hematoxylin and eosin staining of tumors from mice in different groups. Scale bar: 200 μm. (F) Representative images of immunohistochemistry analysis of Ki67 expression in mouse tumor samples. Scale bars, 200 μm. (G) Percentage of Ki67-positive stained cells per field. (H) Immunohistochemical staining for GPX4 and 4-HNE in tissue collected from AN3CA and HEC1B xenografted tumors. Scale bars: 200 μm (Black) or 100 μm (Red). (I) Average optical density of proteins (assessed with ImageJ software). Statistical graphs of immunohistochemistry staining for GPX4 and 4-HNE in tumor xenografts from the indicated groups. * p < 0.05, ** p < 0.01 and *** p < 0.001

Article Snippet: HEC-1B (STCC10605P), AN3CA (STCC10601P), and 293T (STCC10301P) cell lines were purchased from Servicebio (Hubei, China) and were recently authenticated by Short Tandem Repeat profiling.

Techniques: In Vivo, Injection, Negative Control, Saline, Staining, Immunohistochemistry, Expressing, Immunohistochemical staining, Software