lcn2 inhibitor artesunate  (MedChemExpress)

Journal: Clinical and Experimental Medicine

Article Title: The association between 4-HPR-mediated LCN2 suppression and reduced intestinal cell senescence in ulcerative colitis

doi: 10.1007/s10238-025-01843-4

Figure Lengend Snippet: The Drug enrichment analysis. A The result of drug enrichment analysis. B The molecular docking between Fenretinide and LCN2. C The external dataset GSE92415 was utilized to verify the elevated expression of LCN2 in UC (Min to Max, Control, n = 21; UC, n = 162; p -value < 0.0001)

Article Snippet: To delineate the mechanistic pathway, P21 and P16 expression levels were assessed after LPS stimulation in the presence or absence of either the LCN2 inhibitor ZINC00640089 (MedChemExpress) or 4-HPR (MedChemExpress). qRT-PCR was also employed to verify the overexpression of LCN2 in CD4 + cells.

Techniques: Expressing, Control

Journal: Clinical and Experimental Medicine

Article Title: The association between 4-HPR-mediated LCN2 suppression and reduced intestinal cell senescence in ulcerative colitis

doi: 10.1007/s10238-025-01843-4

Figure Lengend Snippet: LCN2 was highly expressed in UC. A P16 was upregulated in Caco-2 cells after the treatment of LPS (n = 3, p -value < 0.0001). P21 was upregulated in Caco-2 cells after the treatment of LPS (n = 3, p -value = 0.0451), LCN2 was upregulated in Caco-2 cells after the treatment of LPS (n = 3, p -value = 0.0005). B The differentially expressed LCN2 between the normal and UC tissues (Symbols and lines, n = 4, p -value = 0.0036). C The expression levels of P16 and P21 following treatment with the LCN2 inhibitor or 4-HPR (n = 3). D – E Colon tissue images and corresponding quantified colon lengths after various treatments (n = 5). F The 7-day body weight change curve (n = 5). G Time-dependent changes in DAI scores following treatment (n = 5). The data of WB analysis was presented as Symbols and lines, the other data are presented as mean ± SEM, * < 0.05, ** < 0.01, *** < 0.001, **** < 0.0001

Article Snippet: To delineate the mechanistic pathway, P21 and P16 expression levels were assessed after LPS stimulation in the presence or absence of either the LCN2 inhibitor ZINC00640089 (MedChemExpress) or 4-HPR (MedChemExpress). qRT-PCR was also employed to verify the overexpression of LCN2 in CD4 + cells.

Techniques: Expressing

Journal: Clinical and Experimental Medicine

Article Title: The association between 4-HPR-mediated LCN2 suppression and reduced intestinal cell senescence in ulcerative colitis

doi: 10.1007/s10238-025-01843-4

Figure Lengend Snippet: 4-HPR attenuated intestinal tissue aging in UC by inhibiting LCN2. A The expression level of senescence-associated genes P16, P21, and LCN2 (n = 5). B The SA-β-gal staining and analysis in colon tissues of three groups (n = 5), scale bar: 200 μm. C Safety assessment of 4-HPR as a single-agent treatment (n = 5). Data are presented as mean ± SEM, **** < 0.0001; ns, no significance

Article Snippet: To delineate the mechanistic pathway, P21 and P16 expression levels were assessed after LPS stimulation in the presence or absence of either the LCN2 inhibitor ZINC00640089 (MedChemExpress) or 4-HPR (MedChemExpress). qRT-PCR was also employed to verify the overexpression of LCN2 in CD4 + cells.

Techniques: Expressing, Staining

Journal: Clinical and Experimental Medicine

Article Title: The association between 4-HPR-mediated LCN2 suppression and reduced intestinal cell senescence in ulcerative colitis

doi: 10.1007/s10238-025-01843-4

Figure Lengend Snippet: 4-HPR regulated the Th17/Treg balance by LCN2. A The levels of inflammatory cytokines of three groups (n = 3). B The gating strategy in flow cytometry. C The distribution of Th17/Treg by flow cytometry analysis (n = 3). D The overexpression of LCN2 was confirmed by qPCR (n = 4). E The secretion level of IL-10 and IL-17A (n = 4). Data are presented as mean ± SEM, * < 0.05, ** < 0.01, *** < 0.001, **** < 0.0001

Article Snippet: To delineate the mechanistic pathway, P21 and P16 expression levels were assessed after LPS stimulation in the presence or absence of either the LCN2 inhibitor ZINC00640089 (MedChemExpress) or 4-HPR (MedChemExpress). qRT-PCR was also employed to verify the overexpression of LCN2 in CD4 + cells.

Techniques: Flow Cytometry, Over Expression

Journal: Cell Death & Disease

Article Title: Lipocalin-2 promotes CKD vascular calcification by aggravating VSMCs ferroptosis through NCOA4/FTH1-mediated ferritinophagy

doi: 10.1038/s41419-024-07260-x

Figure Lengend Snippet: A Representative axial reconstruction of a chest CT scan of a CKD patient without calcification. B Representative sagittal abdominal radiography of a CKD patient without calcification. C Representative axial reconstruction of a chest CT scan of a CKD patient with aortic arch calcification. D Representative sagittal abdominal radiography of a CKD patient with abdominal aorta calcification. The red arrows indicate calcification in the aortic arch and abdominal aorta. E Serum LCN2 levels in patients with CKD with VC (n = 66) or without VC (n = 68) according to abdominal radiography. F Serum LCN2 levels in subgroups defined by AAC scores (n = 105 for mild VC; n = 24 for moderate VC; n = 5 for severe VC). G Scatter diagrams and Spearman coefficient for correlation between serum LCN2 expression level and AAC scores in CKD patients (n = 134). H Scatter diagrams and Spearman coefficient for correlation between the serum LCN2 expression level and the serum calcium level in CKD patients (n = 134). I Serum LCN2 levels in subgroups defined by aortic arch calcium volume (n = 13 for mild VC; n = 7 for moderate VC; n = 6 for severe VC). J Scatter diagrams and Spearman coefficient for correlation between the serum LCN2 expression level and aortic arch calcium volume in CKD patients (n = 26). K Upper: Representative Von Kossa staining images of radial arteries from HD patients with or without VC, scale bar: 50 μm. Middle: Immunofluorescence staining images of LCN2 for Vonkossa staining images, scale bar: 200 μm. Bottom: High-magnification images of the immunofluorescence staining images, scale bar: 50 μm. L Mean fluorescence intensity of LCN2 in the radial arteries of HD patients with VC (n = 12) or without VC (n = 12). M Scatter diagrams and Spearman coefficient for correlation between LCN2 fluorescence intensity and Von Kossa staining-positive areas in the radial arteries of HD patients with or without VC (n = 24). Data are presented as mean ± SD. * p < 0.05, ** p < 0.01 and **** p < 0.0001. VC vascular calcification, CKD chronic kidney disease, CT computed tomography, AAC abdominal aorta calcification score, HD hemodialyzed.

Article Snippet: VSMCs were incubated with LCN2 inhibitor ZINC00784494 (catalogue no. HY-148364, MedChemExpress, New Jersey, USA) under HP conditions (final concentration, 0.5 μmol/L /ml) for 5 days.

Techniques: Computed Tomography, Expressing, Staining, Immunofluorescence, Fluorescence

Journal: Cell Death & Disease

Article Title: Lipocalin-2 promotes CKD vascular calcification by aggravating VSMCs ferroptosis through NCOA4/FTH1-mediated ferritinophagy

doi: 10.1038/s41419-024-07260-x

Figure Lengend Snippet: Basal characteristics of CKD patients with or without VC according to the AAC score.

Article Snippet: VSMCs were incubated with LCN2 inhibitor ZINC00784494 (catalogue no. HY-148364, MedChemExpress, New Jersey, USA) under HP conditions (final concentration, 0.5 μmol/L /ml) for 5 days.

Techniques:

Journal: Cell Death & Disease

Article Title: Lipocalin-2 promotes CKD vascular calcification by aggravating VSMCs ferroptosis through NCOA4/FTH1-mediated ferritinophagy

doi: 10.1038/s41419-024-07260-x

Figure Lengend Snippet: Basal characteristics of CKD patients with or without VC according to the chest CT.

Article Snippet: VSMCs were incubated with LCN2 inhibitor ZINC00784494 (catalogue no. HY-148364, MedChemExpress, New Jersey, USA) under HP conditions (final concentration, 0.5 μmol/L /ml) for 5 days.

Techniques:

Journal: Cell Death & Disease

Article Title: Lipocalin-2 promotes CKD vascular calcification by aggravating VSMCs ferroptosis through NCOA4/FTH1-mediated ferritinophagy

doi: 10.1038/s41419-024-07260-x

Figure Lengend Snippet: Baseline characteristics of HD patients with or without VC according to Von Kossa staining of the radial arteries.

Article Snippet: VSMCs were incubated with LCN2 inhibitor ZINC00784494 (catalogue no. HY-148364, MedChemExpress, New Jersey, USA) under HP conditions (final concentration, 0.5 μmol/L /ml) for 5 days.

Techniques: Staining

Journal: Cell Death & Disease

Article Title: Lipocalin-2 promotes CKD vascular calcification by aggravating VSMCs ferroptosis through NCOA4/FTH1-mediated ferritinophagy

doi: 10.1038/s41419-024-07260-x

Figure Lengend Snippet: A Scheme of the 5/6 nephrectomy and HP diet-induced CKD-VC rat model and AP diet-induced CKD-VC mouse model. B Representative Von Kossa staining and IHC images of LCN2 in the aortic smooth muscle layer from CKD-VC model rats and mice. Scale bar: 200 μm for Von Kossa staining, 100 μm for IHC images. C Quantification of the LCN2-positive area in CKD-VC model rats (n = 6 for the CTL group; n = 8 for the CKD-VC model group). D Quantification of the LCN2-positive area in CKD-VC model mice (n = 5 for the CTL group; n = 7 for the CKD-VC model group). E The expression of LCN2 mRNA in the aortas of CTL and CKD-VC rats was analysed by qRT-PCR (n = 5 per group). Mouse VSMCs were treated with growth medium (CTL) or calcifying medium containing 3.0 mM HP sodium for 5 days. Calcium deposition in VSMCs was assessed by a calcium concentration assay normalised to the protein concentration (n = 3 per group) ( F ) and alizarin red staining ( G ) (positive staining: red; scale bar: 200 μm). H – K The protein levels of osteogenic phenotype markers (RUNX2, BMP2) and LCN2 were determined via Western blot (n = 3 per group). L LCN2 mRNA expression in VSMCs exposed to HP (3.0 mM) (n = 6 per group). Data are presented as mean ± SD. * p < 0.05, ** p < 0.01, *** p < 0.001 and **** p < 0.0001. CKD-VC chronic kidney disease-related vascular calcification, HP high phosphate, VSMCs vascular smooth muscle cells, IHC immunohistochemical, AP adenine and phosphate, CTL control, qRT-PCR quantitative real-time polymerase chain reaction, RUNX2 runt-related transcription factor 2, BMP2 bone morphogenetic protein 2, SD standard deviation.

Article Snippet: VSMCs were incubated with LCN2 inhibitor ZINC00784494 (catalogue no. HY-148364, MedChemExpress, New Jersey, USA) under HP conditions (final concentration, 0.5 μmol/L /ml) for 5 days.

Techniques: Staining, Expressing, Quantitative RT-PCR, Concentration Assay, Protein Concentration, Western Blot, Immunohistochemical staining, Control, Real-time Polymerase Chain Reaction, Standard Deviation

Journal: Cell Death & Disease

Article Title: Lipocalin-2 promotes CKD vascular calcification by aggravating VSMCs ferroptosis through NCOA4/FTH1-mediated ferritinophagy

doi: 10.1038/s41419-024-07260-x

Figure Lengend Snippet: A Scheme of the experiment. After 2 weeks of acclimatisation, LCN2KO and WT mice were fed with a high adenine and phosphate diet (AP) or a normal diet (ND) for 14 weeks. B Survival curves of the experimental mice. Body weights ( C ), serum creatinine ( D ), BUN ( E ), serum phosphorus ( F ) and serum calcium levels ( G ) of the four groups (n = 5 per group). H Western-blot analysis and quantification ( I ) of LCN2 in the aorta (n = 5 per group). J Calcium concentrations of aortas (n = 5 per group). K Representative alizarin red stained images of the aorta. Scale bar:1 cm. L Representative Von Kossa staining of the aortic smooth muscle layer in the four groups and semi-quantification ( M ). Scale bar: 200 μm for 40×; 100 μm for 100×; 50 μm for 200×, 20 μm for 400×. Data are presented as mean ± SD. * p < 0.05, ** p < 0.01, *** p < 0.001 and **** p < 0.0001. CKD-VC chronic kidney disease-related vascular calcification, AP adenine and phosphate, KO knockout, WT wild type, BUN blood urea nitrogen, SD standard deviation.

Article Snippet: VSMCs were incubated with LCN2 inhibitor ZINC00784494 (catalogue no. HY-148364, MedChemExpress, New Jersey, USA) under HP conditions (final concentration, 0.5 μmol/L /ml) for 5 days.

Techniques: Western Blot, Staining, Knock-Out, Standard Deviation

Journal: Cell Death & Disease

Article Title: Lipocalin-2 promotes CKD vascular calcification by aggravating VSMCs ferroptosis through NCOA4/FTH1-mediated ferritinophagy

doi: 10.1038/s41419-024-07260-x

Figure Lengend Snippet: A – C VSMCs were transfected with LCN2 siRNA (siLCN2) or NC siRNA (siNC). The efficacy of transfection was verified by Western blot (n = 4 per group) and qRT-PCR (n = 3 per group). D Calcium deposition in VSMCs was assessed by Alizarin red staining (positive staining: red; scale bar: 200 μm) and quantified using hexadecylpyridinium chloride at an OD of 560 nm ( E ) (n = 6 per group). F Calcium deposition in VSMCs was assessed by calcium concentration assay and was normalised to the protein concentration (n = 3 per group). G Western blot analysis and quantification of the RUNX2 ( H ) and BMP2 ( I ) protein expression in siNC or siLCN2-transfected VSMCs exposed to HP (3.0 mM) or growth medium (CTL) (n = 3 per group). Data are presented as mean ± SD. * p < 0.05, ** p < 0.01, *** p < 0.001 and **** p < 0.0001. qRT-PCR quantitative real-time polymerase chain reaction, HP high phosphate, VSMC vascular smooth muscle cell, siRNA small interfering RNA, NC negative control, OD optical density, CTL control, RUNX2 runt-related transcription factor 2, BMP2 bone morphogenetic protein 2, SD standard deviation.

Article Snippet: VSMCs were incubated with LCN2 inhibitor ZINC00784494 (catalogue no. HY-148364, MedChemExpress, New Jersey, USA) under HP conditions (final concentration, 0.5 μmol/L /ml) for 5 days.

Techniques: Transfection, Western Blot, Quantitative RT-PCR, Staining, Concentration Assay, Protein Concentration, Expressing, Real-time Polymerase Chain Reaction, Small Interfering RNA, Negative Control, Control, Standard Deviation

Journal: Cell Death & Disease

Article Title: Lipocalin-2 promotes CKD vascular calcification by aggravating VSMCs ferroptosis through NCOA4/FTH1-mediated ferritinophagy

doi: 10.1038/s41419-024-07260-x

Figure Lengend Snippet: A Scheme of the experiment. Mice were injected with AAV9-SM22α-vector and AAV9-SM22α- LCN2 and then provided with an AP diet or normal diet (ND) for 14 weeks. B Upper and middle: representative images of Von Kossa staining of the aortic smooth muscle layer. Scale bar: 200 μm for upper; 100 μm for middle; bottom: IHC images of LCN2 expression in four groups. Scale bar: 100 μm. C Semi-quantification of positive Von Kossa staining of the aortic smooth muscle layer D Calcium concentrations of the aorta (n = 5 per group). E Serum phosphorus levels (n = 5 per group). F , G LCN2 overexpression efficiency was validated by Western blot analysis (n = 4 per group). H Calcium deposition in LCN2-overexpressing VSMCs in growth medium or HP conditions was assessed by alizarin red staining (positive staining: red; scale bar: 200 μm) and quantified by hexadecylpyridinium chloride (n = 4 per group) ( I ). J Representative images of alizarin red staining of VSMCs treated with different doses of rLCN2 (0, 250, 500, or 1000 ng/ml) in the presence of HP. Scale bar: 50 μm. K Quantification of the alizarin red staining by hexadecylpyridinium chloride (n = 5 per group). Data are presented as mean ± SD. * p < 0.05, ** p < 0.01, *** p < 0.001 and **** p < 0.0001. AAV adeno-associated virus, Lv lentivirus, CKD-VC chronic kidney disease-related vascular calcification, AP adenine and phosphate, ND normal diet, IHC immunohistochemical, VSMCs vascular smooth muscle cells, HP high phosphate, rLCN2 recombinant LCN2, SD standard deviation.

Article Snippet: VSMCs were incubated with LCN2 inhibitor ZINC00784494 (catalogue no. HY-148364, MedChemExpress, New Jersey, USA) under HP conditions (final concentration, 0.5 μmol/L /ml) for 5 days.

Techniques: Injection, Plasmid Preparation, Staining, Expressing, Over Expression, Western Blot, Virus, Immunohistochemical staining, Recombinant, Standard Deviation

Journal: Cell Death & Disease

Article Title: Lipocalin-2 promotes CKD vascular calcification by aggravating VSMCs ferroptosis through NCOA4/FTH1-mediated ferritinophagy

doi: 10.1038/s41419-024-07260-x

Figure Lengend Snippet: A Representative mRNA expression of PTGS2 in LCN2-knockdown VSMCs under HP or growth medium conditions (n = 5 per group). B Representative mRNA expression of GPX4 in LCN2-knockdown VSMCs under HP or growth medium conditions (n = 5 per group). C Lipid ROS levels were evaluated by C11-BODIPY 581/591 staining. Scale bar: 20 µm. ( D ) MDA content in the aorta. (n = 6 per group). E Typical features of ferroptosis-related changes in the mitochondrial morphology of aortic tissues from mice. (yellow arrows: mitochondria; scale bar:1 µm). F Representative fluorescence images and mean fluorescence intensity ( G ) of the iron probe FerroOrange in LCN2-knockdown VSMCs in the absence or presence of HP (n = 10 per group). Scale bar: 20 µm. H Ferrous iron concentrations in LCN2-knockdown VSMCs in the absence or presence of HP (n = 5 per group). I Molecular docking of the LCN2 protein and NCOA4 protein was performed with clu-spro2.0. The blue colour represents the structure of LCN2, and the red colour represents the molecular structure of NCOA4. J , K VSMCs lysates were immunoprecipitated with anti-LCN2 or anti-NCOA4 antibodies and then analysed by immunoblotting with anti-LCN2 and anti-NCOA4 antibodies. L Representative Western blot images and quantification of NCOA4 ( M ) and FTH1 ( N ) protein expression levels in LCN2- knockdown VSMCs in the absence or presence of HP (n = 4 per group). O Representative Western blot images and quantification of LCN2 ( P ), NCOA4 ( Q ) and FTH1 protein expression ( R ) in the recombinant LCN2-treated VSMCs in the presence or absence of HP (n = 4 per group). Data are presented as mean ± SD. * p < 0.05, ** p < 0.01, *** p < 0.001 and **** p < 0.0001. CKD-VC chronic kidney disease-related vascular calcification, VSMC vascular smooth muscle cell, HP high phosphate, GPX4 glutathione peroxidase 4, ROS reactive oxygen species, MDA malondialdehyde, SD standard deviation.

Article Snippet: VSMCs were incubated with LCN2 inhibitor ZINC00784494 (catalogue no. HY-148364, MedChemExpress, New Jersey, USA) under HP conditions (final concentration, 0.5 μmol/L /ml) for 5 days.

Techniques: Expressing, Knockdown, Staining, Fluorescence, Immunoprecipitation, Western Blot, Recombinant, Standard Deviation

Journal: International Journal of Molecular Sciences

Article Title: Targeting Lipocalin-2 in Inflammatory Breast Cancer Cells with Small Interference RNA and Small Molecule Inhibitors

doi: 10.3390/ijms22168581

Figure Lengend Snippet: siRNA-based silencing in LCN2-overexpressing IBC cells. ( A ) Western blot analysis for LCN2 and β-actin (as loading control) in a panel of IBC (SUM149, SUM190, and MDA-IBC3) and non-IBC (MDA-MB-231, SKBR3, and MCF7) cells. ( B ) Densitometric analysis of band intensities was performed, and values were calculated relative to non-IBC cells, MCF7. Results are shown as Mean ± SEM of triplicate experiments, **** p < 0.001). Two different siRNAs targeting exon 3 and exon 5 of the human LCN2 sequence (NC_000009.12) were used. Western blot analysis of ( C ) MDA-IBC3 cells and ( D ) SUM149 cells were performed after transiently transfected with LCN2-siRNA-1, LCN2-siRNA-2, and negative control-siRNA (NC-siRNA) at 100 nmol/L concentration, as described in materials and methods. Non-treated (NT) cells were treated with the transfection reagent. Densitometric analysis of band intensities of ( E ) MDA-IBC3 and ( F ) SUM149 cells was calculated relative to the NC-siRNA. Results are shown as Mean ± SEM of triplicate experiments (** p < 0.01).

Article Snippet: Twenty-four hours later, LCN2 inhibitors bought to Asinex corporation (North chestnut, NC, USA) were dissolved in DMSO (0.2%) and added to the cells at 10 μM, 1 μM, and 0.1 μM (final concentrations).

Techniques: Western Blot, Control, Sequencing, Transfection, Negative Control, Concentration Assay

Journal: International Journal of Molecular Sciences

Article Title: Targeting Lipocalin-2 in Inflammatory Breast Cancer Cells with Small Interference RNA and Small Molecule Inhibitors

doi: 10.3390/ijms22168581

Figure Lengend Snippet: LCN2-siRNA-based silencing inhibits colony formation, migration, and invasion of IBC cells. Colony formation assay was performed after LCN2-siRNA-based silencing in MDA-IBC3 and SUM149 cells. Cell proliferation was performed in ( A ) MDA-IBC3 cells and ( B ) SUM149 cells. Results are shown as Mean ± SEM of triplicate experiments (* p < 0.05, ** p < 0.01, *** p < 0.001). ( C ) Migration assay was performed after LCN2-siRNA transfection (100 nM siRNA, final concentration) in SUM149. ( D ) NC-siRNA cells represent 100% migration. Images of migrated cells were taken at 20× magnification, scale bar = 100 µm. Results are shown as Mean ± SEM of triplicate experiments (**** p < 0.0001). ( E ) Invasion assay was performed after LCN2-siRNA transfection (100 nM siRNA, final concentration) in SUM149 cells. ( F ) NC-siRNA cells represent 100% invasion. Images of invaded cells were acquired with a light microscope 20× magnification, Scale bar = 100 µm. Results are shown as Mean ± SEM of triplicate experiments (**** p < 0.0001).

Article Snippet: Twenty-four hours later, LCN2 inhibitors bought to Asinex corporation (North chestnut, NC, USA) were dissolved in DMSO (0.2%) and added to the cells at 10 μM, 1 μM, and 0.1 μM (final concentrations).

Techniques: Migration, Colony Assay, Transfection, Concentration Assay, Invasion Assay, Light Microscopy

Journal: International Journal of Molecular Sciences

Article Title: Targeting Lipocalin-2 in Inflammatory Breast Cancer Cells with Small Interference RNA and Small Molecule Inhibitors

doi: 10.3390/ijms22168581

Figure Lengend Snippet: LCN2-siRNA-based silencing induces apoptosis and cell cycle arrest in IBC cells. SUM149 cells were transfected with 100 mM of negative control (NC-siRNA) or LCN2 siRNA (siRNA-2). ( A ) Caspase-3 fluorometric activity assay in SUM149 cells 72 h after LCN2-siRNA-2 and NC-siRNA transfection. Docetaxel (0.5 nM final concentration) was used as a positive control. ( B ) Western blot analysis of apoptotic-related proteins. ( C ) Histogram showing cell cycle arrest at G0/G1 to S phase transition after LCN2-siRNA-2 transfection in SUM149 cells compared with NC-siRNA. ( D ) Quantification of the flow cytometry data showed an increase in SUM149-LCN2-siRNA-2 transfected cells at G0/G1 to S phase transition. ( E ) Western blot analysis of cell cycle-related proteins 72 h after siRNAs transfection. ( F , G ) Densitometric analysis of the band intensities showed in E. Results are shown as Mean ± SEM of triplicate experiments (* p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001).

Article Snippet: Twenty-four hours later, LCN2 inhibitors bought to Asinex corporation (North chestnut, NC, USA) were dissolved in DMSO (0.2%) and added to the cells at 10 μM, 1 μM, and 0.1 μM (final concentrations).

Techniques: Transfection, Negative Control, Activity Assay, Concentration Assay, Positive Control, Western Blot, Sublimation, Flow Cytometry

Journal: International Journal of Molecular Sciences

Article Title: Targeting Lipocalin-2 in Inflammatory Breast Cancer Cells with Small Interference RNA and Small Molecule Inhibitors

doi: 10.3390/ijms22168581

Figure Lengend Snippet: Molecular model and docking of ZINC00784494 and ZINC00640089 ligands into LCN2-calyx pocket. ( A ) Surface model representation of LCN2-calyx pockets. Pockets #1, #2, and #3 (dotted circles) are represented with key amino acid residues in yellow color (right panel). ( B ) Cartoon docking representation and predicted binding interactions of ligands with key residues of LCN2-calyx pocket. Interactions are represented with yellow dotted lines. Residues are displayed with a three-letter code and numbers representing the position in the polypeptide. ( C ) Surface docking representation of ligands (represented as sticks) ZINC00784494 (magenta) and ZINC00640089 (yellow) into the LCN2-calyx pocket.

Article Snippet: Twenty-four hours later, LCN2 inhibitors bought to Asinex corporation (North chestnut, NC, USA) were dissolved in DMSO (0.2%) and added to the cells at 10 μM, 1 μM, and 0.1 μM (final concentrations).

Techniques: Binding Assay

Journal: International Journal of Molecular Sciences

Article Title: Targeting Lipocalin-2 in Inflammatory Breast Cancer Cells with Small Interference RNA and Small Molecule Inhibitors

doi: 10.3390/ijms22168581

Figure Lengend Snippet: LCN2 inhibitors reduce cell proliferation and cell viability in IBC cells. ( A ) For colony formation assays SUM149 cells were treated with LCN2 inhibitors at 10 µM, 1 µM, and 0.1 µM concentration. ( B ) The percentage of clonogenicity was calculated relative to DMSO. Results are shown as Mean ± SEM of triplicate experiments (** p < 0.01, *** p < 0.001). ( C ) Representative plate showing a colony formation assay of SUM149 cells treated with the LCN2 inhibitor. ( D ) Cell viability was assessed in SUM149 cells with Alamar Blue dye 72 h after LCN2 inhibitor treatment. The percentage of cell viability was calculated relative to DMSO. Results are shown as Mean ± SEM of triplicate experiments (**** p < 0.0001).

Article Snippet: Twenty-four hours later, LCN2 inhibitors bought to Asinex corporation (North chestnut, NC, USA) were dissolved in DMSO (0.2%) and added to the cells at 10 μM, 1 μM, and 0.1 μM (final concentrations).

Techniques: Concentration Assay, Colony Assay

Journal: International Journal of Molecular Sciences

Article Title: Targeting Lipocalin-2 in Inflammatory Breast Cancer Cells with Small Interference RNA and Small Molecule Inhibitors

doi: 10.3390/ijms22168581

Figure Lengend Snippet: LCN2 inhibitors ZINC00784494 and ZINC00640089 reduced p-Akt in a dose-dependent manner in SUM149 cells. SUM149 cells were incubated with each inhibitor as described in the “materials and methods” section. Changes in AKT and p-AKT protein levels were measured by Western blot with specific antibodies against these protein forms. ( A ) ZINC00784494, ( B ) ZINC00640089.

Article Snippet: Twenty-four hours later, LCN2 inhibitors bought to Asinex corporation (North chestnut, NC, USA) were dissolved in DMSO (0.2%) and added to the cells at 10 μM, 1 μM, and 0.1 μM (final concentrations).

Techniques: Incubation, Western Blot

Journal: International Journal of Molecular Sciences

Article Title: Targeting Lipocalin-2 in Inflammatory Breast Cancer Cells with Small Interference RNA and Small Molecule Inhibitors

doi: 10.3390/ijms22168581

Figure Lengend Snippet: LCN2-inhibitors ZINC00784494 and ZINC00640089 showed specificity toward LCN2-calyx. Cell proliferation in MCF7, MCF7-EV and, MCF7-LCN2 cells after treatment with ( A ) ZINC00784494 inhibitor and ( B ) ZINC00640089 inhibitor. ( C ) A representative clonogenic assay of MCF7, MCF7-EV, and MCF7-LCN2 treated with ZINC00784494 and ZINC00640089. Results are shown as Mean ± SEM of triplicate experiments (** p < 0.01, *** p < 0.001, **** p < 0.0001).

Article Snippet: Twenty-four hours later, LCN2 inhibitors bought to Asinex corporation (North chestnut, NC, USA) were dissolved in DMSO (0.2%) and added to the cells at 10 μM, 1 μM, and 0.1 μM (final concentrations).

Techniques: Clonogenic Assay