Journal: Journal of Pharmaceutical Analysis

Article Title: Development of a novel NanoBRET high-throughput drug screening assay for human GnRH receptor using sulfo-cyanine 5 fluorophore

doi: 10.1016/j.jpha.2025.101532

Figure Lengend Snippet: Receptor binding assays of sulfo-cyanine 5-D-Lys 6 -gonadotropin-releasing hormone (sCy 5 -D-Lys 6 -GnRH) in SCL60 cells (a human embryonic kidney 293 cell line stably expressing rat gonadotropin-releasing hormone (GnRH) receptor). (A) Saturation binding assay . SCL60 cells were incubated with various concentrations (0, 0.1–100 nM) of sCy 5 -D-Lys 6 -GnRH for 4 h at 4 °C. Following incubation, unbound ligands were removed by washing the cells with ice-cold Dulbecco's phosphate-buffered saline (DPBS), and relative fluorescence intensity/units (RFU) were measured at emission 680 nm (Em680) under excitation of 640 nm (Ex640). Total binding (TB) was assessed in the absence of unlabeled GnRH I, while non-specific binding (NSB) was determined in the presence of a 100-fold excess of unlabeled GnRH I. Specific binding (SB) was calculated by subtracting NSB from TB. (B) Inhibition binding assay. SCL60 cells were incubated with 10 nM sCy 5 -D-Lys 6 -GnRH and increasing concentrations of unlabeled GnRH I (0–10 μM) for 4 h at 4 °C. After incubation, unbound ligands were washed away with ice-cold DPBS, followed by measurement of RFU. Representative results from three independent experiments, each performed in quadruplicate, are shown. Data are presented as mean ± standard deviations (SD).

Article Snippet: Human embryonic kidney 293 (HEK293, ECACC: 85120602) and African green monkey kidney fibroblast (COS-7, ECACC: 87021302) cell lines were purchased from the American Type Culture Collection (Manassas, VA, USA).

Techniques: Binding Assay, Stable Transfection, Expressing, Saturation Assay, Incubation, Saline, Fluorescence, Inhibition

Journal: Journal of Pharmaceutical Analysis

Article Title: Development of a novel NanoBRET high-throughput drug screening assay for human GnRH receptor using sulfo-cyanine 5 fluorophore

doi: 10.1016/j.jpha.2025.101532

Figure Lengend Snippet: Phosphorylation of extracellular signal-regulated kinase 1/2 (p-ERK1/2) induced by sulfo-cyanine 5-D-Lys 6 -gonadotropin-releasing hormone (sCy 5 -D-Lys 6 -GnRH) , gonadotropin-releasing hormone (GnRH) I, and sCy 5 -kisspeptin 10 (sCy 5 -KP10) in human embryonic kidney 293 (HEK293) cell lines stably expressing rat GnRH receptor (SCL60) or human GnRH receptor (hGnRHR). (A) Time-dependent p-ERK1/2. SCL60 cells were treated with 10 nM sCy 5 -D-Lys 6 -GnRH or GnRH I for the indicated times. (B) Dose-dependent p-ERK1/2. SCL60 cells were treated with vehicle (–∞) or increasing concentrations of sCy 5 -D-Lys 6 -GnRH or GnRH I for 5 min. (C) Ligand-induced ERK1/2 phosphorylation and its inhibition by GnRH antagonist Cetrorelix. HEK 293 cells stably expressing hGnRHR were treated with 10 nM of sCy 5 -D-Lys 6 -GnRH, GnRH I, or 1 μM of sCy 5 -KP10, in the absence or presence of 1 μM of Cetrorelix, for 5 min. β-actin was used as a loading control. Representative blots from three independent experiments are presented. ∗∗ P < 0.01, ∗∗∗ P < 0.001 compared with vehicle-treated controls.

Article Snippet: Human embryonic kidney 293 (HEK293, ECACC: 85120602) and African green monkey kidney fibroblast (COS-7, ECACC: 87021302) cell lines were purchased from the American Type Culture Collection (Manassas, VA, USA).

Techniques: Phospho-proteomics, Stable Transfection, Expressing, Inhibition, Control

Journal: Journal of Pharmaceutical Analysis

Article Title: Development of a novel NanoBRET high-throughput drug screening assay for human GnRH receptor using sulfo-cyanine 5 fluorophore

doi: 10.1016/j.jpha.2025.101532

Figure Lengend Snippet: Protein expression levels of N-terminal secretory signal peptide-NanoLuciferase-human gonadotropin-releasing hormone receptor (N-secNluc-hGnRHR) and its mutants. (A) The protein expression levels of N-secNluc-hGnRHR-K191E and N-secNluc-hGnRHR-K191 deletion (K191Δ) mutants relative to the N-secNluc-hGnRHR were determined by the measurement of relative luminescence intensity/units (RLU) in human embryonic kidney 293 (HEK293) cells transiently expressing N-secNluc-hGnRHRs. (B) The cell surface receptor expression levels of N-secNLuc-hGnRHR and N-secNluc-hGnRHR-K191Δ, transiently expressed in African green monkey kidney fibroblast (COS-7) cells, were measured by relative fluorescence intensity/units (RFU) at emission 680 (Em680) nm under excitation of 640 (Ex640) nm through receptor binding assays using 10, 50, and 100 nM, respectively, of sulfo-cyanine 5-D-Lys 6 -gonadotropin-releasing hormone (sCy 5 -D-Lys 6 -GnRH). The N-secNluc-hGnRHR-K191Δ (solid/black bars) construct gave a 4.5 to 5-fold higher receptor expression than that of N-secNluc-hGnRHR (open/white bars) on the cell membrane ( ∗∗∗ P < 0.001, n = 3).

Article Snippet: Human embryonic kidney 293 (HEK293, ECACC: 85120602) and African green monkey kidney fibroblast (COS-7, ECACC: 87021302) cell lines were purchased from the American Type Culture Collection (Manassas, VA, USA).

Techniques: Expressing, Cell Surface Receptor Assay, Fluorescence, Binding Assay, Construct, Membrane

Journal: Journal of Pharmaceutical Analysis

Article Title: Development of a novel NanoBRET high-throughput drug screening assay for human GnRH receptor using sulfo-cyanine 5 fluorophore

doi: 10.1016/j.jpha.2025.101532

Figure Lengend Snippet: Confocal microscopy image analysis of sulfo-cyanine 5-D-Lys 6 -gonadotropin-releasing hormone (sCy 5 -D-Lys 6 -GnRH) binding to human embryonic kidney 293 (HEK293) cells expressing gonadotropin-releasing hormone (GnRH) receptor. (A) Fluorescence of sCy 5 -D-Lys 6 -GnRH bound HEK293 cells expressing N-terminal secretory signal peptide-NanoLuciferase-human gonadotropin-releasing hormone receptor with K191 deletion (N-secNluc-hGnRHR-K191Δ). (B) Overlay of both fluorescence and brightfield/differential interference contrast (DIC) images of the cells in sCy 5 -D-Lys 6 -GnRH bound HEK293 cells expressing N-secNluc-hGnRHR-K191Δ. (C) Abolishment of fluorescence of sCy 5 -D-Lys 6 -GnRH bound HEK293 cells expressing N-secNluc-hGnRHR-K191Δ by co-treatment of the cells with 1 μM of GnRH I. (D) Overlay of both fluorescence and brightfield/DIC images of the HEK293 cells expressing N-secNluc-hGnRHR-K191Δ treated with both sCy 5 -D-Lys 6 -GnRH and 1 μM of GnRH I. (E) The cell membrane localisation of fluorescence of sCy 5 -D-Lys 6 -GnRH bound HEK293 cells expressing GnRHR and the monomeric teal fluorescent protein-H-Ras membrane biomarker (mTFP-H-Ras). (F) Cell membrane localisation of mTFP-H-Ras membrane biomarker of the sCy 5 -D-Lys 6 -GnRH incubated HEK293 cells expressing GnRHR and mTFP-H-Ras.

Article Snippet: Human embryonic kidney 293 (HEK293, ECACC: 85120602) and African green monkey kidney fibroblast (COS-7, ECACC: 87021302) cell lines were purchased from the American Type Culture Collection (Manassas, VA, USA).

Techniques: Confocal Microscopy, Binding Assay, Expressing, Fluorescence, Membrane, Biomarker Discovery, Incubation

Journal: Journal of Pharmaceutical Analysis

Article Title: Development of a novel NanoBRET high-throughput drug screening assay for human GnRH receptor using sulfo-cyanine 5 fluorophore

doi: 10.1016/j.jpha.2025.101532

Figure Lengend Snippet: NanoLuciferase (Nluc) bioluminescence resonance energy transfer (NanoBRET)-based saturation and inhibition binding assays between sulfo-cyanine 5-D-Lys 6 -gonadotropin-releasing hormone (sCy 5 -D-Lys 6 -GnRH) andN-terminal secretory signal peptide-NanoLuciferase-human gonadotropin-releasing hormone receptor and its K191 deletion (N-secNluc-hGnRHR and N-secNluc-hGnRHR-K191Δ) transiently expressed in human embryonic kidney 293 (HEK293) cells. For saturation binding assays, transfected HEK293 cells were incubated with various concentrations (0, 1 nM–100 nM) of sCy 5 -D-Lys 6 -GnRH in the absence of (total binding, TB) or presence (non-specific binding (NSB)) of a 100-fold excess of unlabeled gonadotropin-releasing hormone I (GnRH I) for 30 min at 37 °C, followed by the immediate addition of the substrate furimazine (1:1000) before measurement of emissions. For inhibition assays, transfected HEK293 cells were incubated with 10 nM sCy 5 -D-Lys 6 -GnRH and increasing concentrations of unlabeled GnRH I (0, 0.1 nM to 1 μM) for 4 h at 4 °C. The plates were then brought to room temperature, and furimazine (1:1000) was added immediately before measurement. (A) Saturation binding of sCy 5 -D-Lys 6 -GnRH to N-secNluc-hGnRHR. (B) Saturation binding of sCy 5 -D-Lys 6 -GnRH to N-secNluc-hGnRHR-K191Δ. (C) Inhibition binding of N-secNluc-hGnRHR by GnRH I. (D) Inhibition binding of N-secNluc-hGnRHR-K191Δ by GnRH I. Raw bioluminescence resonance energy transfer (BRET) ratios were calculated by the division of the emission at 610 nm longpass (610LP) by the emission at 460 nm with a bandpass of 80 (460BP80). Representative results from at least three independent experiments, each performed in triplicate, are shown.

Article Snippet: Human embryonic kidney 293 (HEK293, ECACC: 85120602) and African green monkey kidney fibroblast (COS-7, ECACC: 87021302) cell lines were purchased from the American Type Culture Collection (Manassas, VA, USA).

Techniques: Bioluminescence Resonance Energy Transfer, Inhibition, Binding Assay, Transfection, Incubation

Journal: Journal of Pharmaceutical Analysis

Article Title: Development of a novel NanoBRET high-throughput drug screening assay for human GnRH receptor using sulfo-cyanine 5 fluorophore

doi: 10.1016/j.jpha.2025.101532

Figure Lengend Snippet: Kinetics of sulfo-cyanine 5-D-Lys 6 -gonadotropin-releasing hormone (sCy 5 -D-Lys 6 -GnRH) binding toN-terminal secretory signal peptide-NanoLuciferase-human gonadotropin-releasing hormone receptor with K191 deletion (N-secNluc-hGnRHR-K191Δ) stably expressed in human embryonic kidney 293 (HEK293) cells were examined by measurement of bioluminescence resonance energy transfer (BRET) within a continuous time. The cells were seeded into a 96-well plate at a density of 5 × 10 5 cells per well and incubated with furimazine (1:1000 dilution) for 3 min. After the incubation, three different concentrations of sCy 5 -D-Lys 6 -GnRH, approximating the dissociation equilibrium constant ( K D ) value, were added. Emissions were simultaneously measured at 610 nm longpass (610LP) and 460 nm with a bandpass of 80 nm (460BP80) over 40 min at 37 °C. A representative result from three independent experiments, each performed in triplicate, is shown. Data are presented as mean ± standard deviations (SD).

Article Snippet: Human embryonic kidney 293 (HEK293, ECACC: 85120602) and African green monkey kidney fibroblast (COS-7, ECACC: 87021302) cell lines were purchased from the American Type Culture Collection (Manassas, VA, USA).

Techniques: Binding Assay, Stable Transfection, Bioluminescence Resonance Energy Transfer, Incubation

Journal: Journal of Pharmaceutical Analysis

Article Title: Development of a novel NanoBRET high-throughput drug screening assay for human GnRH receptor using sulfo-cyanine 5 fluorophore

doi: 10.1016/j.jpha.2025.101532

Figure Lengend Snippet: NanoLuciferase (Nluc) bioluminescence resonance energy transfer (NanoBRET)-based ligand binding and the Z′ factor of ligand displacements of sulfo-cyanine 5-D-Lys 6 -gonadotropin-releasing hormone (sCy 5 -D-Lys 6 -GnRH) at(N-terminal secretory signal peptide-NanoLuciferase-human gonadotropin-releasing hormone receptor with K191 deletion (N-secNluc-hGnRHR-K191Δ) stably expressed in human embryonic kidney 293 (HEK293) cells. (A) Saturation binding of sCy 5 -D-Lys 6 -GnRH at HEK293 cells stably expressing N-secNluc-hGnRHR-K191Δ. The cells were incubated with the indicated concentrations of sCy 5 -D-Lys 6 -GnRH in the absence of (total binding, TB) or presence (non-specific binding (NSB)) of a 100-fold excess of unlabeled gonadotropin-releasing hormone (GnRH) I for 30 min at 37 °C. The emissions at 610 nm longpass (610LP) and 460 nm with a bandpass of 80 nm (460BP80) were measured following the addition of the substrate furimazine. (B) Competitive inhibition binding of HEK293 cells stably expressing N-secNluc-hGnRHR-K191Δ. The cells were incubated with 10 nM of sCy 5 -D-Lys 6 -GnRH and increasing concentrations of unlabeled Cetrorelix, GnRH I, or GnRH II, ranging from 0 (B 0 , the maximum binding) to 1 μM, and the emissions were then measured as above. (C) Z′ factor values were calculated for the competitive inhibition bindings of sCy 5 -D-Lys 6 -GnRH to N-secNluc-hGnRHR-K191Δ. Data are presented as mean ± standard deviations (SD) from three independent assays performed in triplicate.

Article Snippet: Human embryonic kidney 293 (HEK293, ECACC: 85120602) and African green monkey kidney fibroblast (COS-7, ECACC: 87021302) cell lines were purchased from the American Type Culture Collection (Manassas, VA, USA).

Techniques: Bioluminescence Resonance Energy Transfer, Ligand Binding Assay, Stable Transfection, Binding Assay, Expressing, Incubation, Inhibition

Journal: Journal of Pharmaceutical Analysis

Article Title: Development of a novel NanoBRET high-throughput drug screening assay for human GnRH receptor using sulfo-cyanine 5 fluorophore

doi: 10.1016/j.jpha.2025.101532

Figure Lengend Snippet: Validation of high-throughput screening (HTS) using a small-scale library. Human embryonic kidney 293 (HEK293) cells stably expressing N-terminal secretory signal peptide-NanoLuciferase-human gonadotropin-releasing hormone receptor with K191 deletion (N-secNluc-hGnRHR-K191Δ ) were plated at densities of 5 × 10 5 cells per well in 96-well plates and were then incubated with 10 nM of sulfo-cyanine 5-D-Lys 6 -gonadotropin-releasing hormone (sCy 5 -D-Lys 6 -GnRH) and increasing concentrations of various ligands, such as gonadotropin-releasing hormone (GnRH) I and II, N-lactoyl-phenylalanine, respectively, ranging from 0 (B 0 , the maximum binding) to 10 μM at 4 °C for 4 h. The plates were then brought to room temperature, and furimazine (1:2000, 5 μM) was added and incubated for 5 min. Emissions were simultaneously measured at 610 nm longpass and 460 nm with a bandpass of 80 nm. Raw bioluminescence resonance energy transfer (BRET) ratios were calculated by the division of the emission at 610 nm by the emission at 460 nm. (A) Inhibition of specific BRET signal between sCy 5 -D-Lys 6 -GnRH and N-secNluc-hGnRHR-K191Δ by endogenous GnRHs and their analogues, peptide and non-peptide antagonists. (B) Effect of various GnRH-unrelated molecules on the BRET ratio. Representative results from three independent experiments, each performed in quadruplicate, are shown.

Article Snippet: Human embryonic kidney 293 (HEK293, ECACC: 85120602) and African green monkey kidney fibroblast (COS-7, ECACC: 87021302) cell lines were purchased from the American Type Culture Collection (Manassas, VA, USA).

Techniques: Biomarker Discovery, High Throughput Screening Assay, Stable Transfection, Expressing, Incubation, Binding Assay, Bioluminescence Resonance Energy Transfer, Inhibition, Analogues

Journal: Journal of Pharmaceutical Analysis

Article Title: Development of a novel NanoBRET high-throughput drug screening assay for human GnRH receptor using sulfo-cyanine 5 fluorophore

doi: 10.1016/j.jpha.2025.101532

Figure Lengend Snippet: NanoLuciferase (Nluc) bioluminescence resonance energy transfer (NanoBRET)-based saturation and inhibition binding of Boron-dipyrromethene 630/650-D-Lys 6 -gonadotropin-releasing hormone (BODIPY630/650-D-Lys 6 -GnRH) atN-terminal secretory signal peptide-NanoLuciferase-human gonadotropin-releasing hormone receptor with K191 deletion (N-secNluc-hGnRHR-K191Δ) stably expressed in human embryonic kidney 293 (HEK293) cells. (A) Saturation binding assay. HEK293 cells stably expressing S-N-Nluc-GnRHR-K191Δ were incubated with various concentrations of BODIPY630/650-D-Lys 6 -GnRH in the absence of (total binding, TB) or presence (non-specific binding (NSB)) of a 100-fold excess of unlabeled gonadotropin-releasing hormone (GnRH) I for 30 min at 37 °C. Emissions at 610 nm longpass (610LP) and 460 nm with a bandpass of 80 nm (460BP80) were measured simultaneously following the addition of the substrate furimazine. (B) Inhibition binding assay. HEK293 cells stably expressing S-N-Nluc-GnRHR-K191Δ were incubated with 10 nM BODIPY630/650-D-Lys 6 -GnRH and increasing concentrations of unlabeled GnRH I (0–1 μM) for 4 h at 4 °C. The plate was then brought to room temperature, and furimazine was added immediately before measurement of emissions. The raw bioluminescence resonance energy transfer (BRET) ratios were then calculated. Representative results from at least three independent experiments, each performed in triplicate, are shown.

Article Snippet: Human embryonic kidney 293 (HEK293, ECACC: 85120602) and African green monkey kidney fibroblast (COS-7, ECACC: 87021302) cell lines were purchased from the American Type Culture Collection (Manassas, VA, USA).

Techniques: Bioluminescence Resonance Energy Transfer, Inhibition, Binding Assay, Stable Transfection, Saturation Assay, Expressing, Incubation

Journal: Analytical Chemistry

Article Title: Label-Free Quantitative Thermal Proteome Profiling Reveals Target Transcription Factors with Activities Modulated by MC3R Signaling

doi: 10.1021/acs.analchem.3c03643

Figure Lengend Snippet: Overview of the preparatory and analytical workflows. (A) POMC derived ligands and their downstream signaling cascades. (B) Schematic overview of the thermal proteome profiling (TPP) workflow. MC3R-expressing HEK293 cells were treated with ACTH, α-MSH, or γ-MSH at concentrations of 20, 100, and 500 nM or with DMSO as a vehicle-only negative control. (C) Schematic overview of the TPP data analysis workflow. Protein identification and relative quantification were achieved by direct analysis of the raw LC–MS data, after which various bioinformatics tools were used to infer changes in transcription factor (TF) activity, perform enriched pathway analysis, and identify thermally affected proteins.

Article Snippet: A human embryonic kidney 293 cell line transfected with a tetracycline-regulated expression system to overexpress MC3R (HEK-TREx-MC3R) was provided by Astra Zeneca.

Techniques: Derivative Assay, Expressing, Negative Control, Quantitative Proteomics, Liquid Chromatography with Mass Spectroscopy, Activity Assay

Journal: Analytical Chemistry

Article Title: Label-Free Quantitative Thermal Proteome Profiling Reveals Target Transcription Factors with Activities Modulated by MC3R Signaling

doi: 10.1021/acs.analchem.3c03643

Figure Lengend Snippet: Overview of identified proteins and thermally stabilized or destabilized proteins. (A) Venn diagrams showing the numbers of proteins exhibiting altered melting points, associations with enriched pathways, and phosphorylation in MC3R-expressing HEK293 cells incubated with ACTH, α-MSH, and γ-MSH. (B) Venn diagrams showing the numbers of stabilized, destabilized, and phosphorylated proteins after incubation with ACTH, α-MSH, and γ-MSH. (C) Upset plot representing individual numbers of stabilized and destabilized proteins for each ligand and those common between various combinations of ligands.

Article Snippet: A human embryonic kidney 293 cell line transfected with a tetracycline-regulated expression system to overexpress MC3R (HEK-TREx-MC3R) was provided by Astra Zeneca.

Techniques: Phospho-proteomics, Expressing, Incubation

Journal: Analytical Chemistry

Article Title: Label-Free Quantitative Thermal Proteome Profiling Reveals Target Transcription Factors with Activities Modulated by MC3R Signaling

doi: 10.1021/acs.analchem.3c03643

Figure Lengend Snippet: Characterization of transcription factors. (A) Heat map showing the relative abundance (compared to vehicle-only controls) of the transcription factors CCAR2, HMGB2, DDX21, SRSF7, and TET2 in MC3R-expressing HEK293 cells incubated with ACTH, α-MSH, and γ-MSH at different ligand concentrations and temperatures. (B) Phosphorylation of tryptic peptides derived from the thermally stabilized and destabilized transcription factors shown in panel A whose activity was inferred to change following stimulation with ACTH, α-MSH, or γ-MSH. Phosphorylation sites are indicated by asterisks next to the modified amino acid (shown in parentheses when the exact amino acid is unknown). (C) Transcription factor activities and relational networks inferred from differential expression data using BITFAM. The heatmap shows fold changes in transcription factor activities (relative to vehicle-only treatments) in MC3R-expressing HEK293 cells incubated with ACTH, α-MSH, or γ-MSH. (D) Network showing the interconnectivity of the transcription factors identified within our experimental LC–MS data set.

Article Snippet: A human embryonic kidney 293 cell line transfected with a tetracycline-regulated expression system to overexpress MC3R (HEK-TREx-MC3R) was provided by Astra Zeneca.

Techniques: Expressing, Incubation, Phospho-proteomics, Derivative Assay, Activity Assay, Modification, Quantitative Proteomics, Liquid Chromatography with Mass Spectroscopy

Journal: BMC Medical Genomics

Article Title: VNTR polymorphism in the breakpoint region of ABL1 and susceptibility to bladder cancer

doi: 10.1186/s12920-021-00968-1

Figure Lengend Snippet: Functional analysis of the VNTRs within ABL1 -MS1 in luciferase reporter vector promoter region. a ABL1 promoter vector (p2000) and four promoter vectors in which four different lengths of ABL1 -MS1 (TR13–TR16) were inserted were used. The ABL1 promoter region is indicated by diagonal squares, and the luciferase gene is indicated by gray squares. Four different lengths of ABL1 -MS1 were inserted after the luciferase gene and are indicated by black squares. b The above five different types of promoter vectors were transfected into 293 T and UC3 cells. ABL1 promoter activity was measured by luciferase assay

Article Snippet: The following human cell lines were tested for the effect of ABL1 -MS1 on ABL1 expression: 293 T (human embryonic kidney cell line obtained from the Korean Cell Line Bank; KCLB, Seoul, Korea) and UM-UC3 (bladder cancer cell line).

Techniques: Functional Assay, Luciferase, Plasmid Preparation, Transfection, Activity Assay

Journal: BMC Medical Genomics

Article Title: VNTR polymorphism in the breakpoint region of ABL1 and susceptibility to bladder cancer

doi: 10.1186/s12920-021-00968-1

Figure Lengend Snippet: VNTR analysis of ABL1 -breakpoint cluster region. a Schematic of the VNTR region of ABL1 . 11 exons are marked as black boxes and 10 introns as white boxes. One VNTR region was identified in the ABL1 -breakpoint cluster region, named MS1, and marked with an asterisk. b The location of ABL1 -MS1, the size of the repeating unit, and the consensus sequence were confirmed from the genome information of NCBI

Article Snippet: The following human cell lines were tested for the effect of ABL1 -MS1 on ABL1 expression: 293 T (human embryonic kidney cell line obtained from the Korean Cell Line Bank; KCLB, Seoul, Korea) and UM-UC3 (bladder cancer cell line).

Techniques: Sequencing

Journal: BMC Medical Genomics

Article Title: VNTR polymorphism in the breakpoint region of ABL1 and susceptibility to bladder cancer

doi: 10.1186/s12920-021-00968-1

Figure Lengend Snippet: Allelic type patterns of ABL1 -MS1 in cancer-free male controls and bladder cancer patients. a Haplotype patterns in cancer-free controls and cases with bladder cancer. The left panel is the haplotype pattern for ABL1 -MS1 region from control samples. Three genotypes were identified consisting of two different ABL1 -MS1 alleles. The right panel is the ABL1 -MS1 haplotype pattern seen in bladder cancer patients, showing five genotypes consisting of four different ABL1 -MS1 alleles. The first and last lanes correspond to a 100-bp (M1; Invitrogen Co., CA, USA) and a 1-kb size marker (M2; Invitrogen Co.). b Frequency of genotypes between controls and bladder cancer cases. N corresponds to the total number of samples tested for the allele of ABL1 -MS1. C corresponds to the common alleles (13TR, 15TR) and indicates alleles with a frequency of 1% or more. Rare alleles (R) with a frequency of less than 1% correspond to 14TR and 16TR. * Statistically significant ( P < 0.05)

Article Snippet: The following human cell lines were tested for the effect of ABL1 -MS1 on ABL1 expression: 293 T (human embryonic kidney cell line obtained from the Korean Cell Line Bank; KCLB, Seoul, Korea) and UM-UC3 (bladder cancer cell line).

Techniques: Control, Marker

Journal: BMC Medical Genomics

Article Title: VNTR polymorphism in the breakpoint region of ABL1 and susceptibility to bladder cancer

doi: 10.1186/s12920-021-00968-1

Figure Lengend Snippet: Comparison of allelic frequency of ABL1 -MS1 between controls and bladder cancer

Article Snippet: The following human cell lines were tested for the effect of ABL1 -MS1 on ABL1 expression: 293 T (human embryonic kidney cell line obtained from the Korean Cell Line Bank; KCLB, Seoul, Korea) and UM-UC3 (bladder cancer cell line).

Techniques: Comparison

Journal: BMC Medical Genomics

Article Title: VNTR polymorphism in the breakpoint region of ABL1 and susceptibility to bladder cancer

doi: 10.1186/s12920-021-00968-1

Figure Lengend Snippet: Meiotic segregation of ABL1 -MS1. a Meiotic inheritance of the ABL1 -MS1 in a third-generation family. ABL1 -MS1 were analyzed for minisatellite length in genomic DNA from family members. The pedigree demonstrates the relationship between family groups used in this study: first-generation (lanes 1 and 2, grandfather and grandmother, respectively); second-generation (lanes 3 and 4, father and mother); and third-generation (lanes 5 and 6, children from parents 3 and 4. b Meiotic inheritance the ABL1 -MS1 in three of second-generation. The first-generation is denoted as 1 and 2 (mother and father). The second-generation was shown as 3 and 4, and 5 (children). M corresponds to the size marker

Article Snippet: The following human cell lines were tested for the effect of ABL1 -MS1 on ABL1 expression: 293 T (human embryonic kidney cell line obtained from the Korean Cell Line Bank; KCLB, Seoul, Korea) and UM-UC3 (bladder cancer cell line).

Techniques: Marker

Journal: BMC Medical Genomics

Article Title: VNTR polymorphism in the breakpoint region of ABL1 and susceptibility to bladder cancer

doi: 10.1186/s12920-021-00968-1

Figure Lengend Snippet: Composition of repeats unit of ABL1 -MS1 alleles

Article Snippet: The following human cell lines were tested for the effect of ABL1 -MS1 on ABL1 expression: 293 T (human embryonic kidney cell line obtained from the Korean Cell Line Bank; KCLB, Seoul, Korea) and UM-UC3 (bladder cancer cell line).

Techniques:

Journal: Clinical and Vaccine Immunology : CVI

Article Title: Construction and Immunogenicity of Recombinant Adenovirus Vaccines Expressing the HMW1, HMW2, or Hia Adhesion Protein of Nontypeable Haemophilus influenzae ▿

doi: 10.1128/CVI.00115-10

Figure Lengend Snippet: Bacterial strains, plasmids, and cell lines

Article Snippet: HEK 293 , Human embryonic kidney cell line transfected with the left side of the Ad5 genome , Microbix Biosystems Inc..

Techniques: Plasmid Preparation, Expressing, Derivative Assay, Clone Assay, Transfection