Journal: Oncotarget

Article Title: EFEMP1 promotes ovarian cancer cell growth, invasion and metastasis via activated the AKT pathway

doi: 10.18632/oncotarget.10296

Figure Lengend Snippet: ( A ) Scatter plot showing fold-change values versus P -values for differential gene expression between the highly invasive subclones and the low invasive subclones. The vertical lines correspond to 1.5-fold up and down, respectively, and the horizontal line represents a p -value of 0.05. The red point in the plot represents the differentially expressed genes with statistical significance. ( B , C ) EFEMP1 expression as measured by Western blot (B) and q-RT-PCR (C). ( D ) The fluorescence images of EFEMP1-GFP (Magnification × 200). * P < 0.05 versus control.

Article Snippet: Coverslips were incubated with mouse-anti-human EFEMP1 antibody (sc-33722, Santa Cruz Biotechnology, Inc.) with dilution rate 1:50 at 4°C overnight.

Techniques: Gene Expression, Expressing, Western Blot, Reverse Transcription Polymerase Chain Reaction, Fluorescence, Control

Journal: Oncotarget

Article Title: EFEMP1 promotes ovarian cancer cell growth, invasion and metastasis via activated the AKT pathway

doi: 10.18632/oncotarget.10296

Figure Lengend Snippet: ( A ) EFEMP1 protein expression in lentivirus-infected cells as measured by Western blot. ( B ) EFEMP1 mRNA expression in lentivirus-infected cells as measured by q-RT-PCR. ( C ) PCR products as measured by agarose gel electrophoresis. ( D ) EFEMP1 protein expression in lentivirus-infected cells as measured by ICC staining (Magnification × 200). * P < 0.05 versus control.

Article Snippet: Coverslips were incubated with mouse-anti-human EFEMP1 antibody (sc-33722, Santa Cruz Biotechnology, Inc.) with dilution rate 1:50 at 4°C overnight.

Techniques: Expressing, Infection, Western Blot, Reverse Transcription Polymerase Chain Reaction, Agarose Gel Electrophoresis, Staining, Control

Journal: Oncotarget

Article Title: EFEMP1 promotes ovarian cancer cell growth, invasion and metastasis via activated the AKT pathway

doi: 10.18632/oncotarget.10296

Figure Lengend Snippet: ( A ) Knockdown of EFEMP1 inhibited cell proliferation of the highly invasive subclone. ( B ) Overexpression of EFEMP1 promoted cell proliferation of the low invasive subclone. ( C ) The colony images of lentivirus-infected cells as examined by soft agar colony formation assay. ( D ) EFEMP1 knockdown decreased the colony forming capacity of highly invasive subclone, while overexpression of EFEMP1 increased the colony forming capacity of low invasive subclone. * P < 0.05 versus control.

Article Snippet: Coverslips were incubated with mouse-anti-human EFEMP1 antibody (sc-33722, Santa Cruz Biotechnology, Inc.) with dilution rate 1:50 at 4°C overnight.

Techniques: Knockdown, Over Expression, Infection, Soft Agar Assay, Control

Journal: Oncotarget

Article Title: EFEMP1 promotes ovarian cancer cell growth, invasion and metastasis via activated the AKT pathway

doi: 10.18632/oncotarget.10296

Figure Lengend Snippet: ( A ) Cell cycle analysis as measured by flow cytometry in lentivirus-infected cells. ( B ) EFEMP1 knockdown suppressed cell by blocking their progression from the G1/G0 phase to S phase, while EFEMP1 overexpression promoted cell proliferation by increasing the S phase cells. ( C ) Cell cycle-related proteins Cyclin A, Cyclin B1, Cyclin D1 and Cyclin E in lentivirus-infected cells as measured by Western blot. ( D ) The average band intensities of Cyclin A, Cyclin B1, Cyclin D1 and Cyclin E normalized to GAPDH. * P < 0.05 versus control.

Article Snippet: Coverslips were incubated with mouse-anti-human EFEMP1 antibody (sc-33722, Santa Cruz Biotechnology, Inc.) with dilution rate 1:50 at 4°C overnight.

Techniques: Cell Cycle Assay, Flow Cytometry, Infection, Knockdown, Blocking Assay, Over Expression, Western Blot, Control

Journal: Oncotarget

Article Title: EFEMP1 promotes ovarian cancer cell growth, invasion and metastasis via activated the AKT pathway

doi: 10.18632/oncotarget.10296

Figure Lengend Snippet: ( A ) Cell migration images of lentivirus-infected cells as measured by Boyden chambers without Matrigel. (Magnification × 200). ( B ) Cell invasion images of lentivirus-infected cells by Boyden chambers coated with Matrigel. (Magnification × 200). ( C ) EFEMP1 knockdown suppressed cell migrating abilities, while EFEMP1 overexpression promoted cell migrating abilities. ( D ) EFEMP1 knockdown suppressed cell invading abilities, while EFEMP1 overexpression promoted cell invading abilities. * P < 0.05 versus control.

Article Snippet: Coverslips were incubated with mouse-anti-human EFEMP1 antibody (sc-33722, Santa Cruz Biotechnology, Inc.) with dilution rate 1:50 at 4°C overnight.

Techniques: Migration, Infection, Knockdown, Over Expression, Control

Journal: Oncotarget

Article Title: EFEMP1 promotes ovarian cancer cell growth, invasion and metastasis via activated the AKT pathway

doi: 10.18632/oncotarget.10296

Figure Lengend Snippet: ( A ) EFEMP1 high expressed cells were serum-starved and treated with DMSO or the indicated inhibitors, SB202190, PD98059, and LY294002, for 24 h. The migration and invasion assay were performed by Boyden chambers without or with Matrigel. ( B ) After treating with LY294002, unphosphorylated and phosphorylated forms of AKT were detected by immunoblot analysis. ( C ) The average band intensities were normalized to β-actin. ( D ) After RNA interference, immunoblot analysis were performed to detect the effect of decreased EFEMP1 on the activities of AKT. ( E ) After overexpression transfection, immunoblot analysis were performed to detect the effect of increased EFEMP1 on the activities of AKT. ( F ) The average band intensities were normalized to β-actin. * P < 0.05 versus control.

Article Snippet: Coverslips were incubated with mouse-anti-human EFEMP1 antibody (sc-33722, Santa Cruz Biotechnology, Inc.) with dilution rate 1:50 at 4°C overnight.

Techniques: Migration, Invasion Assay, Western Blot, Over Expression, Transfection, Control

Journal: Oncotarget

Article Title: EFEMP1 promotes ovarian cancer cell growth, invasion and metastasis via activated the AKT pathway

doi: 10.18632/oncotarget.10296

Figure Lengend Snippet: Differential expression of EMT-related genes after EFEMP1 RNA interference

Article Snippet: Coverslips were incubated with mouse-anti-human EFEMP1 antibody (sc-33722, Santa Cruz Biotechnology, Inc.) with dilution rate 1:50 at 4°C overnight.

Techniques: Quantitative Proteomics

Journal: Oncotarget

Article Title: EFEMP1 promotes ovarian cancer cell growth, invasion and metastasis via activated the AKT pathway

doi: 10.18632/oncotarget.10296

Figure Lengend Snippet: ( A ) The microarray image of 84 genes related to tumor progression by RT 2 Profiler™ PCR Array. ( B ) After RNA interference, EMT marker, including E-cadherin, N-cadherin and vimentin as measured by Western blot. ( C ) The top 9 genes (> 5 folds) of the microarray as measure by real-time qPCR in the EFEMP1 shRNA infected group. ( D ) After overexpression transfection, EMT marker, including E-cadherin, N-cadherin and vimentin as measured by Western blot. ( E ) The top 9 genes (> 5 folds) of the microarray as measure by real-time qPCR in the S21-exEFEMP1 infected group. ( F ) The average band intensities were normalized to GAPDH. * P < 0.05 versus control.

Article Snippet: Coverslips were incubated with mouse-anti-human EFEMP1 antibody (sc-33722, Santa Cruz Biotechnology, Inc.) with dilution rate 1:50 at 4°C overnight.

Techniques: Microarray, Marker, Western Blot, shRNA, Infection, Over Expression, Transfection, Control

Journal: Translational Oncology

Article Title: Circulating exosomal protein EFEMP1 and SERPINC1 as diagnostic biomarkers for epithelial ovarian cancer

doi: 10.1016/j.tranon.2024.102126

Figure Lengend Snippet: Characteristics of EOC patients for differential exosomal EFEMP1 and SERPINC1.

Article Snippet: The quantitative detection of the exosomal proteins EFEMP1 and SERPINC1 was performed by the ELISA kits (Boster, Hubei, China) according to the manufacturer's instructions.

Techniques:

Journal: Translational Oncology

Article Title: Circulating exosomal protein EFEMP1 and SERPINC1 as diagnostic biomarkers for epithelial ovarian cancer

doi: 10.1016/j.tranon.2024.102126

Figure Lengend Snippet: Characteristics of benign diseases for differential exosomal EFEMP1 and SERPINC1.

Article Snippet: The quantitative detection of the exosomal proteins EFEMP1 and SERPINC1 was performed by the ELISA kits (Boster, Hubei, China) according to the manufacturer's instructions.

Techniques:

Journal: Translational Oncology

Article Title: Circulating exosomal protein EFEMP1 and SERPINC1 as diagnostic biomarkers for epithelial ovarian cancer

doi: 10.1016/j.tranon.2024.102126

Figure Lengend Snippet: Exosomal EFEMP1 and SERPINC1 were elevated in EOC significantly. The expression levels of EFEMP1 (A) and SERPINC1 (B) were detected by western blot in EOC patients and healthy donors. (H: healthy donors; C: epithelial ovarian cancer); (C) The concentration of exosomal EFEMP1 and SERPINC1 in EOC patients, healthy donors (HD), and patients with benign ovarian tumor (BT); (C) The concentration of exosomal EFEMP1 and SERPINC1 in early-stage (stage I +II), advanced stage (stage III+IV) EOC patients, and HD; (ns, no significance, **** P < 0.0001.).

Article Snippet: The quantitative detection of the exosomal proteins EFEMP1 and SERPINC1 was performed by the ELISA kits (Boster, Hubei, China) according to the manufacturer's instructions.

Techniques: Expressing, Western Blot, Concentration Assay

Journal: Translational Oncology

Article Title: Circulating exosomal protein EFEMP1 and SERPINC1 as diagnostic biomarkers for epithelial ovarian cancer

doi: 10.1016/j.tranon.2024.102126

Figure Lengend Snippet: Exosomal EFEMP1 and SERPINC1 acted as diagnostic biomarker for EOC. (A) The AUCs of exosomal EFEMP1 (left), SERPINC1 (mid), and their combination (right) in EOC patients relative to healthy donors; (B) The AUCs of EFEMP1 (left), SERPINC1 (mid), and their combination (right) in early stage EOC patients relative to healthy donors. AUC: Areas under the curve.

Article Snippet: The quantitative detection of the exosomal proteins EFEMP1 and SERPINC1 was performed by the ELISA kits (Boster, Hubei, China) according to the manufacturer's instructions.

Techniques: Diagnostic Assay, Biomarker Discovery

Journal: Translational Oncology

Article Title: Circulating exosomal protein EFEMP1 and SERPINC1 as diagnostic biomarkers for epithelial ovarian cancer

doi: 10.1016/j.tranon.2024.102126

Figure Lengend Snippet: Exosomal EFEMP1 and SERPINC1 functioned in the differential diagnosis of EOC from benign ovarian tumors. (A) The AUCs of exosomal EFEMP1 (left), SERPINC1 (mid), and their combination (right) in EOC patients relative to patients with benign ovarian tumor; The AUCs of exosomal EFEMP1 (B) or SERPINC1 (C) combined with CA-125 (left) and HE4 (mid), as well as the three combination (right) in EOC patients relative to benign ovarian tumor patients. (D) The combined AUC of CA-125 and HE4(left); and of additional joint with the two exosomal proteins (right) in EOC patients relative to benign ovarian tumor patients. AUC: Areas under the curve.

Article Snippet: The quantitative detection of the exosomal proteins EFEMP1 and SERPINC1 was performed by the ELISA kits (Boster, Hubei, China) according to the manufacturer's instructions.

Techniques: Biomarker Discovery

Journal: Translational Oncology

Article Title: Circulating exosomal protein EFEMP1 and SERPINC1 as diagnostic biomarkers for epithelial ovarian cancer

doi: 10.1016/j.tranon.2024.102126

Figure Lengend Snippet: Exosomal proteins were corelated with coagulation index and LE-DVT. (A) Expression levels of exosomal EFEMP1 and SERPINC1 in serum and plasma. Correlation between exosomal SERPINC1 and platelet count (B) and FIB (C) in EOC patients. (D) Expression level of exosomal SERPINC1 in LE-DVT positive and negative EOC patients. ns, no significance; ** P < 0.01, **** P < 0.0001. FIB: fibrinogen; LE-DVT: lower extremity deep venous thrombosis).

Article Snippet: The quantitative detection of the exosomal proteins EFEMP1 and SERPINC1 was performed by the ELISA kits (Boster, Hubei, China) according to the manufacturer's instructions.

Techniques: Coagulation, Expressing, Clinical Proteomics

Journal: Oncoscience

Article Title: Anti-EGFR function of EFEMP1 in glioma cells and patient prognosis

doi:

Figure Lengend Snippet: (A) Immunoblotting of EGFR in U251 cells treated by human recombinant (hr) EFEMP1 at various concentrations (ng/ml) and for various times (h). (B) Immunoblotting of EGFR and downstream targets in glioma cell lines (U251, LN229, U87) and primary culture (G43-SA) transduced by lentiviral vectors pTRIPZ-Empty (Vector) or pTRIPZ-EFEMP1, with or without a 3-day induction of transgene expression by doxycyclin (Dox, 1 ug/ml). (C) Growth curves of subcutaneous (s.c.) xenografts of U251(pTRIPZ-EFEMP1) by measured tumor volumes and western blotting of EGFR in s.c. xenografts with or without providing Dox in water fed to nude mice. (D) Kaplan-Meier survival curves of nude mice with i.c. implantation of EFEMP1- or Vector-transduced U87 and G43-SA (100,000 cells/implantation).

Article Snippet: The resin/ bead was washed three times with the cold RIPA or TBS buffer (50 mM Tris-Cl, 150 mM NaCl, pH 7.4) to remove on-specifically bound proteins then eluted in 2 X SDS sample buffer and boiled for 3 min prior to loading onto SDS-PAGE gels for immunoblotting using antibodies for EFEMP1 (1:500, Aviva Systems), FLAG-M2 (1:1,000, Sigma-Aldrich), and EGFR (1:1000, Cell Signaling)

Techniques: Western Blot, Recombinant, Plasmid Preparation, Expressing

Journal: Oncoscience

Article Title: Anti-EGFR function of EFEMP1 in glioma cells and patient prognosis

doi:

Figure Lengend Snippet: (A) Depiction of the pTRIPZ-EFEMP1 construct that co-expresses turboRFP and EFEMP1 (with or without an N -terminal FLAG) via the TRE promoter induced by doxycycline (Dox). ( upper ). (B) Infected glioma cells photographed using an inverted phase-contrast, fluorescence microscope. (C) Immunofluorescence (IF) from antibodies for FLAG and EFEMP1 for ectopic EFEMP1 carrying an N -terminal FLAG tag and endogenous/ectopic EFEMP1.

Article Snippet: The resin/ bead was washed three times with the cold RIPA or TBS buffer (50 mM Tris-Cl, 150 mM NaCl, pH 7.4) to remove on-specifically bound proteins then eluted in 2 X SDS sample buffer and boiled for 3 min prior to loading onto SDS-PAGE gels for immunoblotting using antibodies for EFEMP1 (1:500, Aviva Systems), FLAG-M2 (1:1,000, Sigma-Aldrich), and EGFR (1:1000, Cell Signaling)

Techniques: Construct, Infection, Fluorescence, Microscopy, Immunofluorescence, FLAG-tag

Journal: Oncoscience

Article Title: Anti-EGFR function of EFEMP1 in glioma cells and patient prognosis

doi:

Figure Lengend Snippet: Co-immunoprecipitation (IP) of EFEMP1 and EGFR in parental or EFEMP1-transfected U251 (E-CF and E-NF refers to FLAG tags at the C - and N -terminal ends, respectively), with rabbit anti-EGFR antibody and protein A/G beads (A), or mouse anti FLAG M2 affinity gel (B). Rabbit and mouse IgG agarose resin were used as negative controls.

Article Snippet: The resin/ bead was washed three times with the cold RIPA or TBS buffer (50 mM Tris-Cl, 150 mM NaCl, pH 7.4) to remove on-specifically bound proteins then eluted in 2 X SDS sample buffer and boiled for 3 min prior to loading onto SDS-PAGE gels for immunoblotting using antibodies for EFEMP1 (1:500, Aviva Systems), FLAG-M2 (1:1,000, Sigma-Aldrich), and EGFR (1:1000, Cell Signaling)

Techniques: Immunoprecipitation, Transfection

Journal: Oncoscience

Article Title: Anti-EGFR function of EFEMP1 in glioma cells and patient prognosis

doi:

Figure Lengend Snippet: (A) EGF uptake assay in G43-SA and U251 after a 24-hour treatment with Alexa Fluor 647-EGF. (B) Immunofluorescence of EGFR in U251 after a 24-hour treatment with EGF. (C) Immunoblotting of EGFR in U251 whole cell lysate (tEGFR) and membrane-enriched fraction (mEGFR). (D) Immunoblots (left) and plot of normalized densitometry (right) to show the effect of EFEMP1 (a 3-day induction by Dox) on EGFR activation by a low dose of EGF (20 ng/ml). Cells without Dox at each time point were set to unity.

Article Snippet: The resin/ bead was washed three times with the cold RIPA or TBS buffer (50 mM Tris-Cl, 150 mM NaCl, pH 7.4) to remove on-specifically bound proteins then eluted in 2 X SDS sample buffer and boiled for 3 min prior to loading onto SDS-PAGE gels for immunoblotting using antibodies for EFEMP1 (1:500, Aviva Systems), FLAG-M2 (1:1,000, Sigma-Aldrich), and EGFR (1:1000, Cell Signaling)

Techniques: Immunofluorescence, Western Blot, Membrane, Activation Assay

Journal: Oncoscience

Article Title: Anti-EGFR function of EFEMP1 in glioma cells and patient prognosis

doi:

Figure Lengend Snippet: (A) Relative immunohistochemical intensity of EFEMP1 (red) and EGFR (Black) in 62 gliomas of various grades, according to the World Health Organization (WHO) grading system, normalized to averages of three normal cortical tissues. (B) Representative images of consecutive tissue sections from grade I pilocytic astrocytoma (T2), grade II astrocytoma (T4) and diffuse astrocytoma (T8), grade III anaplastic astrocytoma (T14, T16), normal cortex (N), and grade IV glioblastoma multiforme (T39, T42, T48).

Article Snippet: The resin/ bead was washed three times with the cold RIPA or TBS buffer (50 mM Tris-Cl, 150 mM NaCl, pH 7.4) to remove on-specifically bound proteins then eluted in 2 X SDS sample buffer and boiled for 3 min prior to loading onto SDS-PAGE gels for immunoblotting using antibodies for EFEMP1 (1:500, Aviva Systems), FLAG-M2 (1:1,000, Sigma-Aldrich), and EGFR (1:1000, Cell Signaling)

Techniques: Immunohistochemical staining

Journal: Oncoscience

Article Title: Anti-EGFR function of EFEMP1 in glioma cells and patient prognosis

doi:

Figure Lengend Snippet: Prognostic effect of EGFR and EFEMP1 in malignant gliomas based dichotomized subtype

Article Snippet: The resin/ bead was washed three times with the cold RIPA or TBS buffer (50 mM Tris-Cl, 150 mM NaCl, pH 7.4) to remove on-specifically bound proteins then eluted in 2 X SDS sample buffer and boiled for 3 min prior to loading onto SDS-PAGE gels for immunoblotting using antibodies for EFEMP1 (1:500, Aviva Systems), FLAG-M2 (1:1,000, Sigma-Aldrich), and EGFR (1:1000, Cell Signaling)

Techniques:

Journal: Oncoscience

Article Title: Anti-EGFR function of EFEMP1 in glioma cells and patient prognosis

doi:

Figure Lengend Snippet: (A-B) Kaplan-Meier survival curves for two groups of glioma patients dichotomized based on EFEMP1 (cut point 0.06 was for EFEMP1 / ACTB ). Red denotes EGFR -high tumors. Black denotes EGFR -low tumors. (C-D) Kaplan-Meier survival curves for two groups of glioma patients dichotomized based on EGFR (cut point 0.05 was for EGFR / ACTB ). Red denotes EFEMP1 -high tumors. Black denotes EFEMP1 -low tumors.

Article Snippet: The resin/ bead was washed three times with the cold RIPA or TBS buffer (50 mM Tris-Cl, 150 mM NaCl, pH 7.4) to remove on-specifically bound proteins then eluted in 2 X SDS sample buffer and boiled for 3 min prior to loading onto SDS-PAGE gels for immunoblotting using antibodies for EFEMP1 (1:500, Aviva Systems), FLAG-M2 (1:1,000, Sigma-Aldrich), and EGFR (1:1000, Cell Signaling)

Techniques: