Journal: Scientific Reports

Article Title: Development of Potent Forchlorfenuron Analogs and Their Cytotoxic Effect in Cancer Cell Lines

doi: 10.1038/s41598-020-59824-4

Figure Lengend Snippet: The effect of FCF analogs on HER2 expression. ( A ) ECC-1 or HCH-1 cells were incubated with UR214-7 (33 µM), UR214-9 (33 µM) or FCF (300 µM) for 24 h. The levels of each protein were determined by Western blot analysis. ( B ) HCH-1 cells were incubated with UR14-9 (10 µM) or FCF (100 µM) for 48 h (top) or transfected with septin-2 targeting siRNA or non-targeting control siRNA for 24 or 48 h (bottom). Cell lysates were collected. The cellular levels of HER2 were determined by human HER2 enzyme-linked immunosorbent assay. ( C ) The cells were transfected with septin-2 targeting siRNA or non-targeting control siRNA. At 48 h post transfection, cell population and relative expression of septin-2 were determined by sulforhodamine B assay (top) and Western blot analysis (bottom), respectively.

Article Snippet: Following the treatment, cell lysates and supernatants were collected and subjected to HE4 assay (Human HE4 Quantikine ELISA Kit, R&D systems).

Techniques: Expressing, Incubation, Western Blot, Transfection, Control, Enzyme-linked Immunosorbent Assay, Sulforhodamine B Assay

Journal: Scientific Reports

Article Title: Development of Potent Forchlorfenuron Analogs and Their Cytotoxic Effect in Cancer Cell Lines

doi: 10.1038/s41598-020-59824-4

Figure Lengend Snippet: The effect of FCF analogs on the expression of HE4. ( A ) HE4 overexpression clones (OVCAR8-C5) were treated with or without FCF (300 µM) for 7 h, after which the cell lysates and matching culture media were collected and analyzed for the levels of HE4 using enzyme immunoassays. The amount of secreted HE4 was normalized to the protein concentration of respective cell lysate. ( B ) ECC-1 and HCH-1 cells were treated with the indicated concentrations of FCF for 5 h and the secreted levels of HE4 were measured. ( C ) ECC-1 cells were treated with either UR214-1, UR214-7 or FCF at the concentrations indicated for 5 h at which point the secreted levels of HE4 were determined. ( D ) ECC-1 cells were washed and incubated with FCF (300 µM) for 5 h in basal media. Relative HE4 expression was determined by real time PCR (normalized to B2M). ( E ) ECC-1 cells were transfected with septin-2 targeting siRNA or non-targeting control siRNA for 48 h (bottom) or for 72 h (top). Relative gene expression of HE4 was determined by real time PCR (normalized to TBP; bottom). The intracellular levels of HE4 were measured using enzyme immunoassays (top).

Article Snippet: Following the treatment, cell lysates and supernatants were collected and subjected to HE4 assay (Human HE4 Quantikine ELISA Kit, R&D systems).

Techniques: Expressing, Over Expression, Clone Assay, Enzyme Immunoassay, Protein Concentration, Incubation, Real-time Polymerase Chain Reaction, Transfection, Control, Gene Expression

Journal: Oncology Research

Article Title: HE4 Might Participate in Extracellular Matrix Remodeling in Ovarian Cancer via Activation of Fibroblasts

doi: 10.32604/or.2025.069007

Figure Lengend Snippet: The correlation between ovarian tissue stiffness and clinical characteristics. (A) Shore hardness values for normal, benign, and malignant ovarian tissues. (B) Violin plots showing HGSOC tissues hardness compared to both benign ovarian disease and normal tissues. (C) The correlation between serum HE4 levels and ovarian tissue stiffness in EOC (n = 70). (D) The correlation between serum HE4 levels and tissue stiffness in HGSOC patients (n = 46). ** p < 0.01, *** p < 0.001, **** p < 0.0001, ns = not significant

Article Snippet: Subsequently, the proteins were transferred to a polyvinylidene fluoride membrane (Millipore, Burlington, MA, USA) and blocked with 5% skimmed milk at 37°C for 2 h. The membrane was then incubated overnight at 4°C with primary antibodies HE4 (1:1000; Proteintech, 66557-1-Ig, Wuhan, China), α-SMA (1:1000; Proteintech, 14395-1-Ig), FAP (1:800; HUABIO, ET1704-23, Hangzhou, China), Collagen I (1:800; Proteintech, 67288-1-Ig), Vimentin (1:2000; HUABIO, ET1610-39) and GAPDH (1:10,000; Proteintech, 60004-1-Ig).

Techniques:

Journal: Oncology Research

Article Title: HE4 Might Participate in Extracellular Matrix Remodeling in Ovarian Cancer via Activation of Fibroblasts

doi: 10.32604/or.2025.069007

Figure Lengend Snippet: Expression of HE4 levels, fibroblast activation markers and collagen in high vs. low stiffness groups. (A) Serum HE4 levels in the high stiffness group and low stiffness group (n = 11/group). (B) The protein levels of HE4 in different stiffness groups were assayed by Western blot (n = 11/group). (C) Representative IHC staining of HE4 of cancer cell in high and low groups (n = 11/group), Scale bars, 50 μm/20 μm. (D) Western blot results show that the expression of fibroblast activation marker α-SMA (n = 11/group). (E) Immunohistochemistry results demonstrate fibroblast activation markers and Collagen I of mesenchymal areas in high and low groups (n = 11/group), Scale bars, 50 μm/20 μm. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001

Article Snippet: Subsequently, the proteins were transferred to a polyvinylidene fluoride membrane (Millipore, Burlington, MA, USA) and blocked with 5% skimmed milk at 37°C for 2 h. The membrane was then incubated overnight at 4°C with primary antibodies HE4 (1:1000; Proteintech, 66557-1-Ig, Wuhan, China), α-SMA (1:1000; Proteintech, 14395-1-Ig), FAP (1:800; HUABIO, ET1704-23, Hangzhou, China), Collagen I (1:800; Proteintech, 67288-1-Ig), Vimentin (1:2000; HUABIO, ET1610-39) and GAPDH (1:10,000; Proteintech, 60004-1-Ig).

Techniques: Expressing, Activation Assay, Western Blot, Immunohistochemistry, Marker

Journal: Oncology Research

Article Title: HE4 Might Participate in Extracellular Matrix Remodeling in Ovarian Cancer via Activation of Fibroblasts

doi: 10.32604/or.2025.069007

Figure Lengend Snippet: Cell-secreted HE4 induced fibroblast activation and extracellular matrix deposition. (A) HE4 knockdown cell lines were established in OVCAR3/OVCAR8 cells using lentiviral transfection, and the knockdown efficiency was verified by Western blot (n = 3/group). (B) Western blot analysis expression of α-SMA and FAP after NOFs were co-cultured with sh-HE4 or control groups of OVCAR3/OVCAR8 (n = 3/group). (C) The fibroblast markers of α-SMA and vimentin were analyzed by immunohistochemistry OVCAR3(NC: n = 24, sh1: n = 21, sh2: n = 17)/OVCAR8(NC: n = 40, sh1: n = 40, sh2: n = 40). Scale bar, 20 μm (OVCAR3)/10 μm (OVCAR8). (D) Effect of HE4 knockdown on collagen contractility OVCAR3/OVCAR8 (n = 9/group). (E) Treatment of NOFs co-cultured with the sh-HE4 groups led to decreased the level of Collagen I OVCAR3/OVCAR8 (n = 3/group). * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001, ns = not significant

Article Snippet: Subsequently, the proteins were transferred to a polyvinylidene fluoride membrane (Millipore, Burlington, MA, USA) and blocked with 5% skimmed milk at 37°C for 2 h. The membrane was then incubated overnight at 4°C with primary antibodies HE4 (1:1000; Proteintech, 66557-1-Ig, Wuhan, China), α-SMA (1:1000; Proteintech, 14395-1-Ig), FAP (1:800; HUABIO, ET1704-23, Hangzhou, China), Collagen I (1:800; Proteintech, 67288-1-Ig), Vimentin (1:2000; HUABIO, ET1610-39) and GAPDH (1:10,000; Proteintech, 60004-1-Ig).

Techniques: Activation Assay, Knockdown, Transfection, Western Blot, Expressing, Cell Culture, Control, Immunohistochemistry

Journal: Oncology Research

Article Title: HE4 Might Participate in Extracellular Matrix Remodeling in Ovarian Cancer via Activation of Fibroblasts

doi: 10.32604/or.2025.069007

Figure Lengend Snippet: Fibroblast activation and collagen deposition were inhibited in vivo . (A) Representative images of tumors excised from mice injected with Normal Control (NC) or sh-HE4 OVCAR3 cells (n = 5/group). (B) Tumor volume was measured over 21 days, tumors in the HE4 knockdown groups grew significantly slower compared to the control group. (C) Tumor weight at the endpoint. Tumors in the HE4 knockdown groups were significantly lighter than those in the control group. (D) Representative IHC staining of HE4 and quantification of HE4-positive areas in NC and HE4 knockdown groups (n = 5/group). (E) Representative IHC staining of and quantification of fibroblast activation markers and Collagen I positive areas (n = 5/group). * p < 0.05, ** p < 0.01, **** p < 0.0001

Article Snippet: Subsequently, the proteins were transferred to a polyvinylidene fluoride membrane (Millipore, Burlington, MA, USA) and blocked with 5% skimmed milk at 37°C for 2 h. The membrane was then incubated overnight at 4°C with primary antibodies HE4 (1:1000; Proteintech, 66557-1-Ig, Wuhan, China), α-SMA (1:1000; Proteintech, 14395-1-Ig), FAP (1:800; HUABIO, ET1704-23, Hangzhou, China), Collagen I (1:800; Proteintech, 67288-1-Ig), Vimentin (1:2000; HUABIO, ET1610-39) and GAPDH (1:10,000; Proteintech, 60004-1-Ig).

Techniques: Activation Assay, In Vivo, Injection, Control, Knockdown, Immunohistochemistry

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: ZNF703 promotes tumor progression in ovarian cancer by interacting with HE4 and epigenetically regulating PEA15

doi: 10.1186/s13046-020-01770-0

Figure Lengend Snippet: ZNF703 interacted with HE4 and HE4 promoted the nuclear translocation of the ZNF703. a The co-immunoprecipitation analysis of ZNF703 and HE4 was performed in ovarian cancer cells (The left two were performed in CAOV3 cell line and the right two were performed in OVCAR3 cell line. The corresponding molecular weight was added to the right side). b The immunofluorescence co-localization of ZNF703 and HE4 was performed in CAOV3 and OVCAR3 cells (The green color was for ZNF703 and the red color was for HE4, DAPI was blue, the colocalization was yellow) (×800). The arrow indicated the co-localization in the nucleus. c, d Western blot analysis of intracellular total ZNF703 protein in CAOV3 and OVCAR3 cells transfected with the overexpressed HE4 plasmid or control, or HE4 siRNA or control. e, f Western blot determined nuclear and cytoplasmic distribution of ZNF703 after nuclear cytoplasmic fractionation in CAOV3 and OVCAR3 cells transfected with the overexpressed HE4 plasmid or control. g, h Nuclear and cytoplasmic distribution of ZNF703 tested by western blot after nuclear cytoplasmic fractionation in CAOV3 and OVCAR3 cells transfected with HE4 siRNA or control. i, Immunofluorescent staining of ZNF703 in CAOV3 and OVCAR3 cells transfected with HE4 siRNA or control (× 800). The statistical results are on the right side of their corresponding positions. The experiment was repeated three times. Data are presented as mean ± SD. *, P < 0.05; **, P < 0.01; ***, P < 0.001

Article Snippet: Subsequently, the cells were incubated with the primary antibody against ZNF703 (1:30) diluted in blocking buffer at 4 °C overnight and then incubated with Alexa Fluor 488-goat anti-mouse IgG (1:100) secondary antibody for 2 h. Next, the cells were rinsed with PBS for 15 min, and the nuclei were stained using 4′,6-diamidino-2-phenylindole dihydrochloride (DAPI) for 5 min. For detecting co-localization, the anti-HE4 primary antibody (1:200, DF8160, Affinity) and the anti-ZNF703 primary antibody (1:30, Santa Cruz) were incubated at the same time.

Techniques: Translocation Assay, Immunoprecipitation, Molecular Weight, Immunofluorescence, Western Blot, Transfection, Plasmid Preparation, Control, Fractionation, Staining

Journal: Journal of Ovarian Research

Article Title: HE4 promotes collateral resistance to cisplatin and paclitaxel in ovarian cancer cells

doi: 10.1186/s13048-016-0240-0

Figure Lengend Snippet: HE4-overexpressing cells are more resistant to cisplatin and paclitaxel treatment. SKOV3-NV, SKOV3-C1, and SKOV3-C7 cells were treated with 0-12.5 μM cisplatin ( a ) or 0-5 nM paclitaxel ( b ) for 48 h, at which time the cells were subjected to MTS assay to measure viability. Results are displayed as percent survival of vehicle treated cells. Error bars represent the standard deviation of biological replicates. * p < .05, ** p < .005, N/S = not significant ( c ) SKOV3-NV, SKOV3-C1, and SKOV3-C7 cells were treated with vehicle (DMSO) or 5 μM cisplatin, and protein was collected at 24, 48, and 72 h after treatment. Western blot was performed to detect levels of PARP and cleaved PARP. d Densitometry analysis of PARP and cleaved PARP normalized to GAPDH, from western blot in ( c ). e SKOV3-NV, SKOV3-C1, and SKOV3-C7 cells were treated with 1 nM paclitaxel and protein was isolated from all cells (floating and adherent) 48 h after treatment. Western blot was performed to detect levels of PARP and cleaved PARP. GAPDH was used as a loading control. f Densitometry analysis of PARP and cleaved PARP normalized to GAPDH, from western blot in ( e )

Article Snippet: To generate HE4-CRISPR Double Nickase stable cell lines, SKOV3-C1 cells were transfected in 6-well plates with 1 μg HE4 Double Nickase Plasmid (Santa Cruz, sc-402876-NIC) or Control Double Nickase Plasmid (Santa Cruz, sc-437281), using 5 μl Lipofectamine 2000 (Invitrogen).

Techniques: MTS Assay, Standard Deviation, Western Blot, Isolation, Control

Journal: Journal of Ovarian Research

Article Title: HE4 promotes collateral resistance to cisplatin and paclitaxel in ovarian cancer cells

doi: 10.1186/s13048-016-0240-0

Figure Lengend Snippet: CRISPR/Cas-mediated HE4 downregulation reduces chemoresistance in SKOV3-C1 cells. a SKOV3-C1 cells were transfected with a human CRISPR-HE4 Double Nickase plasmid and CRISPR-Control Double Nickase plasmid for 48 h. Transfected cells were selected with puromycin and efficient downregulation of HE4 was measured by qRT-PCR and ELISA. [HE4] = concentration of HE4 in supernatant as measured by ELISA. b - c CRISPR-Control and CRISPR-HE4 cells were treated with DMSO vehicle or 25 μM cisplatin or 5 nM paclitaxel for 24 h, and MTS assay was performed to measure cell viability. Results are displayed as percent survival of vehicle treated cells. Error bars represent the standard deviation of three or more biological replicates, * p < .05

Article Snippet: To generate HE4-CRISPR Double Nickase stable cell lines, SKOV3-C1 cells were transfected in 6-well plates with 1 μg HE4 Double Nickase Plasmid (Santa Cruz, sc-402876-NIC) or Control Double Nickase Plasmid (Santa Cruz, sc-437281), using 5 μl Lipofectamine 2000 (Invitrogen).

Techniques: CRISPR, Transfection, Plasmid Preparation, Control, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Concentration Assay, MTS Assay, Standard Deviation

Journal: Journal of Ovarian Research

Article Title: HE4 promotes collateral resistance to cisplatin and paclitaxel in ovarian cancer cells

doi: 10.1186/s13048-016-0240-0

Figure Lengend Snippet: EGR1 mRNA and protein upregulation is suppressed in HE4-overexpressing clones. a Quantitative RT-PCR validation of microarray using RNA from SKOV3-NV, SKOV3-C1, and SKOV3-C7 cells treated with vehicle (DMSO) or 25 μM cisplatin. Error bars represent standard deviation of three biological replicates, * p < .05, ** p < .005. b Timecourse analysis of EGR1 expression following treatment of SKOV3-NV and SKOV3-C1 cells with 25 μM cisplatin. c EGR1, phosphorylated ERK, and total ERK protein levels following treatment with vehicle (DMSO), 25 μM cisplatin, or 10 ng/mL rEGF (positive control) were determined by western blot. GAPDH was used as a loading control. d - e Densitometry analysis of EGR1 (normalized to GAPDH) and phospho-ERK/ERK ratio, from western blot in ( c )

Article Snippet: To generate HE4-CRISPR Double Nickase stable cell lines, SKOV3-C1 cells were transfected in 6-well plates with 1 μg HE4 Double Nickase Plasmid (Santa Cruz, sc-402876-NIC) or Control Double Nickase Plasmid (Santa Cruz, sc-437281), using 5 μl Lipofectamine 2000 (Invitrogen).

Techniques: Clone Assay, Quantitative RT-PCR, Biomarker Discovery, Microarray, Standard Deviation, Expressing, Positive Control, Western Blot, Control

Journal: Journal of Ovarian Research

Article Title: HE4 promotes collateral resistance to cisplatin and paclitaxel in ovarian cancer cells

doi: 10.1186/s13048-016-0240-0

Figure Lengend Snippet: HE4 promotes deregulation of mitogen activated protein kinase (MAPK) signaling. a Protein was collected from SKOV3-NV, SKOV3-C1, and SKOV3-C7 cells and OVCAR8-NV and OVCAR8-C5 cells and western blot performed for phosphorylated ERK and total ERK. b Densitometry analysis of phospho-ERK/ERK ratio, from western blot in ( a ). c SKOV3-WT and OVCAR8-WT cells were treated with 20 nM recombinant HE4 for the indicated times, or left untreated. Protein was collected and western blot performed to determine levels of phosphorylated ERK and total ERK. d Densitometry analysis of phospho-ERK/ERK ratio, from western blot in ( c ). e SKOV3-NV and SKOV3-C1 cells were treated with vehicle (DMSO) or 25 μM cisplatin for the indicated times and western blot performed to detect levels of phospho-p38 and total p38. f Densitometry analysis of phospho-p38/p38 ratio, from western blot in ( e )

Article Snippet: To generate HE4-CRISPR Double Nickase stable cell lines, SKOV3-C1 cells were transfected in 6-well plates with 1 μg HE4 Double Nickase Plasmid (Santa Cruz, sc-402876-NIC) or Control Double Nickase Plasmid (Santa Cruz, sc-437281), using 5 μl Lipofectamine 2000 (Invitrogen).

Techniques: Western Blot, Recombinant

Journal: Journal of Ovarian Research

Article Title: HE4 promotes collateral resistance to cisplatin and paclitaxel in ovarian cancer cells

doi: 10.1186/s13048-016-0240-0

Figure Lengend Snippet: HE4 promotes tubulin deregulation in SKOV3 and OVCAR8 cells. a Quantitative RT-PCR of MAPT in SKOV3-NV and SKOV3-C1 cells, normalized to 18 s rRNA. ** p < .05 b SKOV3-WT and OVCAR8-WT cells were left untreated or treated with 20 nM recombinant HE4, and protein was collected at the indicated timepoints. Levels of β-tubulin and α-tubulin were determined by western blot, with GAPDH used as a loading control. c - d Densitometry analysis of β-tubulin and α-tubulin, normalized to GAPDH, from western blot in ( b )

Article Snippet: To generate HE4-CRISPR Double Nickase stable cell lines, SKOV3-C1 cells were transfected in 6-well plates with 1 μg HE4 Double Nickase Plasmid (Santa Cruz, sc-402876-NIC) or Control Double Nickase Plasmid (Santa Cruz, sc-437281), using 5 μl Lipofectamine 2000 (Invitrogen).

Techniques: Quantitative RT-PCR, Recombinant, Western Blot, Control

Journal: Scientific reports

Article Title: Recurrence monitoring for ovarian cancer using a cell phone-integrated paper device to measure the ovarian cancer biomarker HE4/CRE ratio in urine.

doi: 10.1038/s41598-021-01544-4

Figure Lengend Snippet: Figure 1. Comparison between flatbed scanner and cell phone measurement of HE4 concentrations (left) and of creatinine concentrations (right). Error bars the percent error found for each concentration (mean + s.d, n = 4). The dotted line indicates the line of identity. Error bars the percent error.

Article Snippet: To identify the best reporter, we screened three monoclonal HE4 reporter antibodies (R&D systems, LsBio, and Hytest).

Techniques: Comparison, Concentration Assay

Journal: Scientific reports

Article Title: Recurrence monitoring for ovarian cancer using a cell phone-integrated paper device to measure the ovarian cancer biomarker HE4/CRE ratio in urine.

doi: 10.1038/s41598-021-01544-4

Figure Lengend Snippet: Figure 3. Comparison of mean values for HE4/CRE ratios for scanner and cellphone to actual values (mean + s.d, n = 5). The dotted line indicates the line of identity.

Article Snippet: To identify the best reporter, we screened three monoclonal HE4 reporter antibodies (R&D systems, LsBio, and Hytest).

Techniques: Comparison

Journal: Scientific reports

Article Title: Recurrence monitoring for ovarian cancer using a cell phone-integrated paper device to measure the ovarian cancer biomarker HE4/CRE ratio in urine.

doi: 10.1038/s41598-021-01544-4

Figure Lengend Snippet: Figure 2. Simulated clinical patient ratios of urinary HE4/CRE. Each image shows a lateral flow strip and corresponding creatinine strip below. In the LFA strip, the left line is the test line and the right line is the control line. *LMP low malignancy potential cyst. The labels below are the ratio of HE4/CRE and the corresponding stage of ovarian cancer given by Liao et al. (2015) and Hellstrom et al. (2010).

Article Snippet: To identify the best reporter, we screened three monoclonal HE4 reporter antibodies (R&D systems, LsBio, and Hytest).

Techniques: Stripping Membranes, Control

Journal: Scientific reports

Article Title: Recurrence monitoring for ovarian cancer using a cell phone-integrated paper device to measure the ovarian cancer biomarker HE4/CRE ratio in urine.

doi: 10.1038/s41598-021-01544-4

Figure Lengend Snippet: Figure 4. Schematic of the two paper test strips and components. For the HE4 test strip (top), the sandwich assay captures the biomarker in the middle at the test line. The control line is generated when excess reporter binds. For the creatinine test strip, a two reagent system is used to generate a colorimetric reaction with creatinine from light to dark purple. The intensity of the purple color (bottom) indicates the amount of creatinine. The two strips are placed in a scanner or cassette for the cell phone to analyze.

Article Snippet: To identify the best reporter, we screened three monoclonal HE4 reporter antibodies (R&D systems, LsBio, and Hytest).

Techniques: Stripping Membranes, Biomarker Discovery, Control, Generated

Journal: Scientific reports

Article Title: Recurrence monitoring for ovarian cancer using a cell phone-integrated paper device to measure the ovarian cancer biomarker HE4/CRE ratio in urine.

doi: 10.1038/s41598-021-01544-4

Figure Lengend Snippet: Figure 6. Calibration plots generated from the scanner. (A) Standard curve for concentrations of HE4 (pM) (mean + s.d, n = 2). (B) Standard curve for concentrations of creatinine (mg/dL) (mean + s.d, n = 3).

Article Snippet: To identify the best reporter, we screened three monoclonal HE4 reporter antibodies (R&D systems, LsBio, and Hytest).

Techniques: Generated

Journal: Scientific reports

Article Title: Recurrence monitoring for ovarian cancer using a cell phone-integrated paper device to measure the ovarian cancer biomarker HE4/CRE ratio in urine.

doi: 10.1038/s41598-021-01544-4

Figure Lengend Snippet: Figure 5. HE4 test line intensity as a function of the assay read time (mean + s.d., n = 2) (left panel). The color intensity as a function of assay read time for three creatinine concentrations of 5 mg/dL, 20 mg/dL, and 40 mg/ dL (mean + s.d, n = 3) (right panel).

Article Snippet: To identify the best reporter, we screened three monoclonal HE4 reporter antibodies (R&D systems, LsBio, and Hytest).

Techniques: Functional Assay

Journal: Scientific reports

Article Title: Recurrence monitoring for ovarian cancer using a cell phone-integrated paper device to measure the ovarian cancer biomarker HE4/CRE ratio in urine.

doi: 10.1038/s41598-021-01544-4

Figure Lengend Snippet: Figure 7. Overview of the smartphone quantification system. (A) Image of the prototype components (lens, cell phone attachments and strip cassette). (B) A CAD model providing an overview of the components used in developing the system. (C) Schematic of the test-strip holder indicating the color detection scheme and the red boxes (arrows) indicate the region of interest (ROI) for HE4, for background reference, and a ROI for CRE.

Article Snippet: To identify the best reporter, we screened three monoclonal HE4 reporter antibodies (R&D systems, LsBio, and Hytest).

Techniques: Stripping Membranes

Journal: Scientific reports

Article Title: Recurrence monitoring for ovarian cancer using a cell phone-integrated paper device to measure the ovarian cancer biomarker HE4/CRE ratio in urine.

doi: 10.1038/s41598-021-01544-4

Figure Lengend Snippet: Figure 8. Calibration plots generated from the cell phone-based device. (A) Standard curve for concentrations of HE4 (pM) (mean + s.d, n = 2). (B) Standard curve for concentrations of creatinine (mg/dL) from cell phone (mean + s.d, n = 3).

Article Snippet: To identify the best reporter, we screened three monoclonal HE4 reporter antibodies (R&D systems, LsBio, and Hytest).

Techniques: Generated

Journal: Frontiers in Medicine

Article Title: Integrative Informatics Analysis of Transcriptome and Identification of Interacted Genes in the Glomeruli and Tubules in CKD

doi: 10.3389/fmed.2020.615306

Figure Lengend Snippet: Verification of WFDC2 and PEX19 interaction in HK2, podocyte, and glomerular endothelial cells. (A) Relative WFDC2 transcript levels were upregulated in many diabetic nephropathy human and mice datasets in the kidney tubule. (B) Relative FEX19 transcript levels were downregulated in FSGS in the kidney glomeruli in Nephroseq database. (C,D) WFDC2 in the tubule was upregulated, and FEX19 in the glomeruli was downregulated in STZ-induced diabetic mice by qPCR and western blot. (E) Western blot shows WFDC2 overexpression in HK2 cell. (F) PEX19 protein level was decreased with WFDC2 treatment in podocyte and glomerular endothelial cells by western blot. Con, WFDC2 empty vector; OE, WFDC2 overexpression vector; EC, glomerular endothelial cells. (A–C) Values are mean ± SEM; * p ≤ 0.05 between means. (A,B) All the genes were significantly changed in CKD compared with control with p < 0.001.

Article Snippet: The following antibodies were used: WFDC2 (rabbit monoclonal HE4/WFDC2, Catalog # NBP2-66883; Novus Biologicals), PEX19 (PEX19 monoclonal antibody (GT554), Catalog # MA5-17266; Invitrogen), and GAPDH (mouse monoclonal antibody, Catalog # G8795-100UL; Sigma).

Techniques: Western Blot, Over Expression, Plasmid Preparation, Control