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Image Search Results
Journal: Genomics
Article Title: PLOD1 promote proliferation and migration with glycolysis via the Wnt/β-catenin pathway in THCA.
doi: 10.1016/j.ygeno.2024.110943
Figure Lengend Snippet: Fig. 1. PLOD1 expression was positively correlated with THCA progression. A-C. PLOD1 expression in TCGA-THCA database was analyzed by the Ualcan website. D. The overall survival (OS) rates were determined using GEPIA website. E-G. PLOD1 expression in THCA tissues and its paired adjacent normal tissues was assessed via western blot, qRT-PCR and immunohistochemistry assay. H and I. PLOD1 levels in THCA cells (BHT101, KTC1, TPC1 and CAL-62) and the primary human thyroid follicular epithelial cell line (Nthy-ori 3–1) were tested via qRT-PCR and western blot. *P < 0.05.
Article Snippet: The primary antibodies:
Techniques: Expressing, Western Blot, Quantitative RT-PCR, Immunohistochemistry
Journal: Genomics
Article Title: PLOD1 promote proliferation and migration with glycolysis via the Wnt/β-catenin pathway in THCA.
doi: 10.1016/j.ygeno.2024.110943
Figure Lengend Snippet: Fig. 2. PLOD1 depletion dampened THCA cell proliferation, while enhanced apoptosis. The sequences of si-NC and si-PLOD-1, 2, and 3 were transfected into BHT101 and TPC1 cells, respectively. The vector and pcDNA3.1-PLOD1 was transfected into KTC1 cells. A. Cell transfection efficiency was tested via western blot. B–D. BHT101, TPC1 cell and KTC1 cell viability, colony formation and proliferation were assessed by CCK- 8 assay, colony formation assay and EdU corporation assay, respectively. E. BHT101 and TPC1 cell apoptosis was analyzed via flow cytometry analysis. F. Apoptosis- related proteins (Bcl-2, Bax and cleaved-PARP) expression were assessed by western blot. *P < 0.05, **P < 0.01.
Article Snippet: The primary antibodies:
Techniques: Transfection, Plasmid Preparation, Western Blot, CCK-8 Assay, Colony Assay, Flow Cytometry, Expressing
Journal: Genomics
Article Title: PLOD1 promote proliferation and migration with glycolysis via the Wnt/β-catenin pathway in THCA.
doi: 10.1016/j.ygeno.2024.110943
Figure Lengend Snippet: Fig. 3. PLOD1 depletion impeded THCA cell invasion and migration. The sequences of si-NC and si-PLOD-1 and 3 were transfected into BHT101 and TPC1 cells, respectively. The vector and pcDNA3.1-PLOD1 was transfected into KTC1 cells. A. BHT101, TPC1 and KTC1 cell migration was assessed by wound healing assay. B.EMT-related proteins (E-cadherin, Vimentin and N-cadherin) expression were evaluated by western blot. C and D. BHT101, TPC1 and KTC1 cell migration and invasion were assessed via transwell assay. **P < 0.01.
Article Snippet: The primary antibodies:
Techniques: Migration, Transfection, Plasmid Preparation, Wound Healing Assay, Expressing, Western Blot, Transwell Assay
Journal: Genomics
Article Title: PLOD1 promote proliferation and migration with glycolysis via the Wnt/β-catenin pathway in THCA.
doi: 10.1016/j.ygeno.2024.110943
Figure Lengend Snippet: Fig. 4. PLOD1 might function as an oncogene in THCA via being positively regulated by its transcription factor MAZ. A. The upstream transcription factor of PLOD1 was predicted with the hTFtarget database. The transcription factor positively correlated with PLOD1 that derived from TCGA-THCA was identified by Venn diagram analysis. B. The result of Pearson correlation analysis between PLOD1 and MAZ. C. The targeting relationship between PLOD1 and MAZ was analyzed by dual-luciferase reporter assay. D. The empty vector and recombinant MAZ overexpressing vector were transfected into BHT101 and TPC1 cells, respectively. PLOD1 and MAZ levels were detected by western blot. E and F. BHT101 and TPC1 cell proliferation and apoptosis were tested by colony formation assay and flow cytometry analysis, respectively. G. BHT101 and TPC1 cell migration and invasion were assessed by transwell assay. *P < 0.05.
Article Snippet: The primary antibodies:
Techniques: Derivative Assay, Luciferase, Reporter Assay, Plasmid Preparation, Recombinant, Transfection, Western Blot, Colony Assay, Flow Cytometry, Migration, Transwell Assay
Journal: Genomics
Article Title: PLOD1 promote proliferation and migration with glycolysis via the Wnt/β-catenin pathway in THCA.
doi: 10.1016/j.ygeno.2024.110943
Figure Lengend Snippet: Fig. 5. PLOD1 depletion impeded THCA cell glycolysis. The sequences of si-NC and si-PLOD-1 and 3 were transfected into BHT101 and TPC1 cells, respectively. A and B. GSEA with PLOD1 in TCGA-THCA was carried out with the LinkedOmics webtool. C-E. Glycolysis-related indicators including glucose uptake, lactate production and ATP/ADP ratio were assessed. F. Glycolysis-related proteins expression levels were detected by western blot. *P < 0.05.
Article Snippet: The primary antibodies:
Techniques: Transfection, Expressing, Western Blot
Journal: Genomics
Article Title: PLOD1 promote proliferation and migration with glycolysis via the Wnt/β-catenin pathway in THCA.
doi: 10.1016/j.ygeno.2024.110943
Figure Lengend Snippet: Fig. 6. PLOD1 depletion exerted tumor-impeding roles in THCA via deactivating the Wnt/β-catenin pathway. BHT101 and TPC1 cells were treated with 0.5 μM of BML-284 for 48 h. The sequences of si-NC and si-PLOD-1, 2, and 3 were transfected into BHT101 and TPC1 cells, respectively. A. GSEA was also conducted with the cluster Profiler package in R using the TCGA-THCA database. B. GSEA curve of the Wnt/β-catenin pathway. C. Wnt/β-catenin pathway-related proteins (Cyclin D1, β-catenin, GSK-3β and WNT5A) expression were assessed by western blot. D. The β-catenin expression in the nucleus and cytoplasm was detected by nucleocytoplasmic separation western blot. E. The nuclear localization of β-catenin in BHT101 and TPC1 was detected using immuno fluorescence. F. The TOP/FOP luciferase reporter assay in PLOD1-depleted BHT101 and TPC1 cells. G. Wnt/β-catenin pathway-related proteins (Cyclin D1, β-catenin, GSK-3β and WNT5A) expression were assessed by western blot. H and I. BHT101 and TPC1 cell proliferation and apoptosis were tested by colony formation assay and flow cytometry analysis, respectively. J. BHT101 and TPC1 cell migration and invasion were analyzed by transwell assay. *P < 0.05.
Article Snippet: The primary antibodies:
Techniques: Transfection, Expressing, Western Blot, Fluorescence, Luciferase, Reporter Assay, Colony Assay, Flow Cytometry, Migration, Transwell Assay
Journal: Genomics
Article Title: PLOD1 promote proliferation and migration with glycolysis via the Wnt/β-catenin pathway in THCA.
doi: 10.1016/j.ygeno.2024.110943
Figure Lengend Snippet: Fig. 7. PLOD1 depletion suppressed THCA tumor growth and glycolysis via deactivating the Wnt/β-catenin pathway. The nude mice were divided into the NC group and the PLOD1-depletion group. Each mouse was subcutaneously injected with 100 μl of shRNA-PLOD1 or shRNA-NC transfected TPC1 cells (3.5 × 103 cells/μl) on the dorsal flanks. A-C. The tumor volume and weight of each group were calculated and recorded. D and E. Cell proliferation and apoptosis in the generated tumor were tested by immunohistochemistry assay and TUNEL assay, respectively. F and G. Expression of glycolysis and Wnt/β-catenin pathway-related proteins were tested by western blot. *P < 0.05, **P < 0.01.
Article Snippet: The primary antibodies:
Techniques: Injection, shRNA, Transfection, Generated, Immunohistochemistry, TUNEL Assay, Expressing, Western Blot
Journal: Orphanet Journal of Rare Diseases
Article Title: Visualization of cholesterol deposits in lysosomes of Niemann-Pick type C fibroblasts using recombinant perfringolysin O
doi: 10.1186/1750-1172-9-64
Figure Lengend Snippet: Selective recognition of cholesterol by GST-PFO. (A) Protein-lipid overlay assay. Indicated amounts of lipids were spotted on nitrocellulose membrane and incubated with 1 μg/ml GST-PFO. Immunoreactive spots were revealed by chemiluminescence. (B) 6-Carboxyfluorescein release from liposomes of indicated composition induced by 15 μg/ml GST-PFO. The data are expressed as percentage of the total amount of 6-carboxyfluorescein released from liposomes by 0.2% Triton X-100. (C) SPR sensograms of binding of 10 μg/ml GST-PFO or 20 μg/ml GST to liposomes of indicated lipid composition immobilized on the surface of sensor chip L1. Binding of GST-PFO to liposomes was carried out for 600 s after which the samples were washed for another 600 s. In (A) and (C) data represent one of three independent experiments; in (B) mean ± SEM from three experiments is shown. Chol – cholesterol; Cer – ceramide; SM – sphingomyelin.
Article Snippet: In studies on colocalization of PFO-stained structures, the
Techniques: Protein-lipid Overlay Assay (PLOA), Membrane, Incubation, Liposomes, Binding Assay
Journal: Orphanet Journal of Rare Diseases
Article Title: Visualization of cholesterol deposits in lysosomes of Niemann-Pick type C fibroblasts using recombinant perfringolysin O
doi: 10.1186/1750-1172-9-64
Figure Lengend Snippet: GST-PFO labels cholesterol deposits in NPC fibroblasts. (A-C) NPC fibroblasts were cultured in DMEM/10% FBS (A, A’) or in DMEM supplemented with delipidated 10% FBS for 72 h (B-C’) . Inset in (B’) shows an enlargement of a merged image of an area marked in (B, B’) . Colocalization of GST-PFO and filipin staining is in white. Colocalization of the labels is also seen in vesicles marked by arrows in (B, B’) . (C, C’) Omitting GST-PFO during labeling of permeabilized NPC cells yields only traces of non-specific staining with secondary antibodies (C) . In these conditions, filipin detects cholesterol accumulated in the cells (C’) . (D-D’) Fibroblasts from a healthy donor were cultured in DMEM/delipidated 10% FBS. Cells were fixed, permeabilized with 0.05% Triton X-100 and incubated with 5 μg/ml GST-PFO followed by secondary antibodies conjugated with Texas Red (left) or with 25 μg/ml filipin (right) to compare the staining patterns revealed by the two probes. Scale bar, 20 μm.
Article Snippet: In studies on colocalization of PFO-stained structures, the
Techniques: Cell Culture, Staining, Labeling, Incubation
Journal: Orphanet Journal of Rare Diseases
Article Title: Visualization of cholesterol deposits in lysosomes of Niemann-Pick type C fibroblasts using recombinant perfringolysin O
doi: 10.1186/1750-1172-9-64
Figure Lengend Snippet: GST-PFO labels vesicles containing DiI-LDL in NPC cells. NPC fibroblasts (A-A”) and healthy control fibroblasts (B-B”) were cultured in DMEM supplemented with delipidated 10% FBS for 72 h, incubated with 10 μg/ml DiI-LDL for 5 h and, after fixation and permeabilization, processed for GST-PFO labeling. (A, B) GST-PFO staining, (A’, B’) fluorescence of DiI-LDL, (A”, B”) merged images. Inset in (A” ) shows an enlargement of a marked area. Scale bar, 20 μm.
Article Snippet: In studies on colocalization of PFO-stained structures, the
Techniques: Control, Cell Culture, Incubation, Labeling, Staining, Fluorescence
Journal: Orphanet Journal of Rare Diseases
Article Title: Visualization of cholesterol deposits in lysosomes of Niemann-Pick type C fibroblasts using recombinant perfringolysin O
doi: 10.1186/1750-1172-9-64
Figure Lengend Snippet: Colocalization of GST-PFO with LAMP-1 and filipin in NPC fibroblasts. Cholesterol visualized by GST-PFO (red) is present in lysosomes identified by anti-LAMP-1 (green in A’-A” ) and filipin (blue in B’-B” ). (A”s1-s4) Merged images of optical sections of the cell shown in (A-A”) . The sections range from the basolateral (A”s1) to the apical (A”s4) part of the cell. (A”, B”) Merged images of (A-A’) and (B-B’) , respectively. Colocalization of GST-PFO and LAMP-1 is seen as yellow color in (A” and A”s1-s4) while pink color reflects colocalization of GST-PFO- and filipin-positive structures in (B”) . Scale bars, 20 μm.
Article Snippet: In studies on colocalization of PFO-stained structures, the
Techniques:
Journal: Orphanet Journal of Rare Diseases
Article Title: Visualization of cholesterol deposits in lysosomes of Niemann-Pick type C fibroblasts using recombinant perfringolysin O
doi: 10.1186/1750-1172-9-64
Figure Lengend Snippet: Quantitation of colocalization of GST-PFO-positive structures and other cellular components in NPC fibroblasts. (A) Pearson’s correlation coefficient, (B) overlap coefficient for co-immunostaining with GST-PFO and filipin or antibodies against LAMP1, LBPA, golgin-84, PDI, PMP70 or phalloidin used as an actin filament marker. TX, Triton X-100, DiT, digitonin. Data shown are mean ± SEM.
Article Snippet: In studies on colocalization of PFO-stained structures, the
Techniques: Quantitation Assay, Immunostaining, Marker
Journal: Orphanet Journal of Rare Diseases
Article Title: Visualization of cholesterol deposits in lysosomes of Niemann-Pick type C fibroblasts using recombinant perfringolysin O
doi: 10.1186/1750-1172-9-64
Figure Lengend Snippet: GST-PFO labeling of NPC cells colocalizes with LBPA. NPC (A-B”) and healthy control fibroblasts (C-C”) were fixed, permeabilized with 0.05% Triton X-100 (A-A”) or 0.05% digitonin (B-C”) and labeled with GST-PFO and anti-LBPA antibody. (A, B, C) GST-PFO labeling, (A’, B’, C’) LBPA localization, (A”, B”, C”) merged images. TX, Triton X-100, DiT, digitonin. Scale bar, 20 μm.
Article Snippet: In studies on colocalization of PFO-stained structures, the
Techniques: Labeling, Control
Journal: Orphanet Journal of Rare Diseases
Article Title: Visualization of cholesterol deposits in lysosomes of Niemann-Pick type C fibroblasts using recombinant perfringolysin O
doi: 10.1186/1750-1172-9-64
Figure Lengend Snippet: Negligible co-immunostaining of GST-PFO-positive structures and other organelles in NPC cells. (A-D) Distribution of GST-PFO-positive structures, (A’-D’) patterns of staining by antibodies directed against golgin-84 (A’) , PDI - an endoplasmic reticulum marker (B’) , anti-PMP70 – a peroxisomal marker (C’) and phalloidin labeling actin filaments (D’) . (A”-D”) merging of the corresponding pairs of images displays traces of the yellow color indicating different localization of GST-PFO-positive structures and the examined proteins. Scale bar, 20 μm.
Article Snippet: In studies on colocalization of PFO-stained structures, the
Techniques: Immunostaining, Staining, Marker, Labeling
Journal: Orphanet Journal of Rare Diseases
Article Title: Visualization of cholesterol deposits in lysosomes of Niemann-Pick type C fibroblasts using recombinant perfringolysin O
doi: 10.1186/1750-1172-9-64
Figure Lengend Snippet: Immunoelectron microscopy analysis of GST-PFO staining in NPC and healthy fibroblasts. Ultrathin sections of NPC cells (A, B) or control healthy fibroblasts (C) were subjected to post-embedding immunogold labeling with GST-PFO followed by anti-GST-biotin and anti-biotin-gold antibodies (6 nm) together with rabbit anti-LAMP-1 and anti-rabbit-gold antibodies (10 nm). GST-PFO decorates material accumulated in vesicles of NPC cells (arrows in A , B ) while LAMP-1 is visualized mainly at the borders of vesicles in NPC and control fibroblasts (semicircles in A and C ).
Article Snippet: In studies on colocalization of PFO-stained structures, the
Techniques: Immuno-Electron Microscopy, Staining, Control, Labeling
Journal: Orphanet Journal of Rare Diseases
Article Title: Visualization of cholesterol deposits in lysosomes of Niemann-Pick type C fibroblasts using recombinant perfringolysin O
doi: 10.1186/1750-1172-9-64
Figure Lengend Snippet: Determination of cholesterol content in NPC cells by cellular ELISA. (A) Binding of GST-PFO to cholesterol increases proportionally to the number of cells in the sample within the range of 1-15×10 3 cells/well. Triton X-100-permeabilized NPC fibroblasts were exposed to 5 μg/ml GST-PFO followed by anti-GST IgY-peroxidase. Absorbance of the product of peroxidase activity is shown. (B) Permeabilization of cells with Triton X-100 does not extract cholesterol from deposits. Fixed NPC cells were treated with 0.03%-0.2% Triton-X-100 (TX) or 0.05% digitonin, as indicated, and processed for cellular ELISA using GST-PFO. In a series of experiments, prior to labeling with GST-PFO, cells were incubated with 3 mM methyl-β (CDX) for 60 min at 37°C. In these conditions prominent reduction in the binding of GST-PFO to cells is observed. (C) Significant binding of GST-PFO to NPC fibroblasts. NPC cells and healthy controls were processed for cellular ELISA. When indicated, prior to labeling with GST-PFO cells were incubated with 3 mM methyl-β-cyclodextrin. The results are presented as a ratio of absorbance at 450 nm and 595 nm. Data are means ± SEM from four experiments. (D-D’) In non-permeabilized cells cholesterol deposits are not stained with GST-PFO (D) but are labeled with filipin (D’) . (D”) : merged images of D and D’ . Scale bar, 20 μm.
Article Snippet: In studies on colocalization of PFO-stained structures, the
Techniques: Enzyme-linked Immunosorbent Assay, Binding Assay, Activity Assay, Labeling, Incubation, Staining