Journal: Clinical and Translational Medicine

Article Title: Elevation of ISG15 promotes diabetic kidney disease by modulating renal tubular epithelial cell pyroptosis

doi: 10.1002/ctm2.70337

Figure Lengend Snippet: ISG15 was involved in HG‐induced mitochondrial impairment and mtDNA release. (A) Representative TEM images of kidney tissues from WT and KO mice treated with STZ ( n = 6). (B) Western blot analysis ISG15/ISGylation expression in TECs treated with vehicle or HG ( n = 3). (C–E) Flow cytometry analysis and quantitative data depicting the mitochondrial membrane potential (C), mitochondrial mass (D) and mtROS (E) ( n = 3). (F and G) qPCR analysis the mtDNA ( Loop1‐3 and mt‐Nd4 ) copy number in the cytosolic compartments (F) and mitochondria (G) ( n = 3). TECs were treated with vehicle or C‐176 (10 µM), and then cultured in HG medium for 48 h. Results are expressed as the mean ± SD. * p < .05; ** p < .01; *** p < .001; ns, not significant.

Article Snippet: To inhibit mitochondrial reactive oxygen species (mtROS), NLRP3 or STING, cells were pretreated with the mitochondria‐specific antioxidant Mito‐TEMPO (25 μM; TargetMol, China), NLRP3‐specific inhibitor MCC950 (2 μM; TargetMol) or STING‐specific inhibitor C‐176 (10 μM; TargetMol) for 1.5 h before exposure to 40 mM HG.

Techniques: Western Blot, Expressing, Flow Cytometry, Membrane, Cell Culture

Journal: Disease Markers

Article Title: circIFT80 Functions as a ceRNA for miR-142, miR-568, and miR-634 and Promotes the Progression of Colorectal Cancer by Targeting β -Catenin

doi: 10.1155/2022/8081246

Figure Lengend Snippet: circIFT80 was in abnormally high expression in CRC tissues and cells. (a) circIFT80 was highly expressed in CRC tissues compared with adjacent normal tissue, as determined by microarray analysis. (b) Quantitative PCR (qPCR) analysis of the relative expression levels of circIFT80 in normal tissues and CRC tissues; paired t -test; ∗∗ P < 0.01. (c) Quantitative PCR (qPCR) analysis of the circIFT80 relative expression level in both CRC cells (HT-29 and SW480) and normal colonic cell (HcoEpic). ∗∗ P < 0.01.

Article Snippet: CCK-8 solution was used to detect the viability of CRC cells and was purchased from TargetMol (Boston, USA).

Techniques: Expressing, Microarray, Real-time Polymerase Chain Reaction

Journal: Disease Markers

Article Title: circIFT80 Functions as a ceRNA for miR-142, miR-568, and miR-634 and Promotes the Progression of Colorectal Cancer by Targeting β -Catenin

doi: 10.1155/2022/8081246

Figure Lengend Snippet: circIFT80 overexpression facilitated the proliferation and colony formation of CRC cells. (a) Cell viability of CRC cells as detected by CCK-8 assays following transfection with si-circIFT80 or OE circIFT80 into HT-29 cells. (b) Cell viability of CRC cells was detected by CCK-8 assays transfection with si-circIFT80 or OE circIFT80 into SW480 cells. (c) Colony formation assays were conducted in CRC cells transfected with si-circIFT80 or Ov-circIFT80 into HT-29 cells. (d) Colony formation assays were conducted in CRC cells transfected with si-circIFT80 or Ov-circIFT80 into SW480 cells. ∗ P < 0.05 and ∗∗ P < 0.01. si-circIFT80: circIFT80 silencing; si-NC: the control sequence of si-circIFT80; Ov-circIFT80: overexpressed circIFT80; NC: pcDNA3.1 plasmid.

Article Snippet: CCK-8 solution was used to detect the viability of CRC cells and was purchased from TargetMol (Boston, USA).

Techniques: Over Expression, CCK-8 Assay, Transfection, Control, Sequencing, Plasmid Preparation

Journal: Disease Markers

Article Title: circIFT80 Functions as a ceRNA for miR-142, miR-568, and miR-634 and Promotes the Progression of Colorectal Cancer by Targeting β -Catenin

doi: 10.1155/2022/8081246

Figure Lengend Snippet: circIFT80 facilitated the ability of CRC cells to migrate. (a, b) Wound healing assays in HT-29 cells transfected with si-circIFT80 or Ov-circIFT80. Quantitative data is shown below. (c, d) Wound healing assays in SW480 cells transfected with si-circIFT80 or Ov-circIFT80. Quantitative data is shown below. Quantitative data is shown below. (e, f) The protein level of MMP2 and MMP9 was measured by western blot in HT-29 cells with si-circIFT80 or Ov-circIFT80. (g, h) MMP2 and MMP9 expression was detected by a western blot in SW480 cells with si-circIFT80 or Ov-circIFT80. ∗ P < 0.05 and ∗∗ P < 0.01. The data from three independent experiments.

Article Snippet: CCK-8 solution was used to detect the viability of CRC cells and was purchased from TargetMol (Boston, USA).

Techniques: Transfection, Western Blot, Expressing

Journal: Disease Markers

Article Title: circIFT80 Functions as a ceRNA for miR-142, miR-568, and miR-634 and Promotes the Progression of Colorectal Cancer by Targeting β -Catenin

doi: 10.1155/2022/8081246

Figure Lengend Snippet: circIFT80 contributed to cell cycle progression and reduced cellular apoptosis. (a) The extent of apoptosis in CRC cells was detected by flow cytometry following the transfection of si-NC and si-circIFT80. (b) The extent of apoptosis in CRC cells was detected by flow cytometry following transfection with Ov-circIFT80 or NC. Quantitative results are shown in the left panel. (c) The cell cycle distribution of CRC cells was analyzed by flow cytometry. Quantitative data are shown below. ∗ P < 0.05 and ∗∗ P < 0.01.

Article Snippet: CCK-8 solution was used to detect the viability of CRC cells and was purchased from TargetMol (Boston, USA).

Techniques: Flow Cytometry, Transfection

Journal: Disease Markers

Article Title: circIFT80 Functions as a ceRNA for miR-142, miR-568, and miR-634 and Promotes the Progression of Colorectal Cancer by Targeting β -Catenin

doi: 10.1155/2022/8081246

Figure Lengend Snippet: β -Catenin expression was regulated by circIFT80 in CRC cells. (a) The expression levels of circIFT80 were analyzed by qPCR in HT-29 cells transfected with si-NC, si-circIFT80, pcDNA3.1, and pcDNA-circIFT80. (b) The expression levels of β -catenin (CTNNB1) were measured by qPCR in HT-29 cells transfected with si-NC, si-circIFT80, pcDNA3.1, and pcDNA-circIFT80. (c) The protein levels of β -catenin and c-myc were measured by western blotting in HT-29 cells transfected with si-NC, si-circIFT80, pcDNA3.1, and pcDNA-circIFT80. Quantitative data are shown in the right panel. GAPDG was used as a loading control. ∗∗ P < 0.01 and ∗∗∗ P < 0.001.

Article Snippet: CCK-8 solution was used to detect the viability of CRC cells and was purchased from TargetMol (Boston, USA).

Techniques: Expressing, Transfection, Western Blot, Control

Journal: Disease Markers

Article Title: circIFT80 Functions as a ceRNA for miR-142, miR-568, and miR-634 and Promotes the Progression of Colorectal Cancer by Targeting β -Catenin

doi: 10.1155/2022/8081246

Figure Lengend Snippet: circIFT80 acted as a molecular sponge for miR-142, miR-568, and miR-634 in CRC cells. (a) circIFT80 containing wild-type (WT) or mutant (MUT) binding sites along with the sequence complementarity between miR-142, miR-634, and miR-568. (b) Relative luciferase activity was detected in 293 T cells after cotransfection with the WT or MUT circIFT80 reporter plasmids in NC or miR-142, miR-634, and miR-568 mimic. (c) circIFT80 expression levels were analyzed by qPCR after RNA pull-down assays using biotin-labeled miR-142, miR-568, and miR-634 probes and a control probe. (d) In situ hybridization assay of the colocation of circIFT80 and miR-142, miR-568, and miR-634. (e) The expression level of miR-142, miR-568, and miR-634 between CRC cells (HT-29 and SW480) and normal human colonic epithelial cells (HcoEpic) was analyzed by qPCR assay. (f) The expression level of miR-142, miR-568, and miR-634 between CRC tissues and tumor adjacent normal tissues was detected by qPCR assay. (g) The cell viability of HT-29 cells following different transfections, as evaluated by CCK-8 assays. U6 was used as an internal control. ∗ P < 0.05 and ∗∗ P < 0.01.

Article Snippet: CCK-8 solution was used to detect the viability of CRC cells and was purchased from TargetMol (Boston, USA).

Techniques: Mutagenesis, Binding Assay, Sequencing, Luciferase, Activity Assay, Cotransfection, Expressing, Labeling, Control, In Situ Hybridization, Transfection, CCK-8 Assay

Journal: Journal of Cancer Research and Clinical Oncology

Article Title: Identified γ-glutamyl cyclotransferase (GGCT) as a novel regulator in the progression and immunotherapy of pancreatic ductal adenocarcinoma through multi-omics analysis and experiments

doi: 10.1007/s00432-024-05789-0

Figure Lengend Snippet: GGCT hinders the advancement of pancreatic cancer by inducing the upregulation of c-Myc. A GSEA enrichment analysis of hallmark gene sets for GGCT in multiple pancreatic cancer datasets. B GSEA analysis of GO enrichment for GGCT. C GSEA analysis of KEGG enrichmen for GGCT. D GSEA analysis of c-Myc releted gene set for GGCT. E Correlation analysis of GGCT and c-Myc in PDAC. F Western blot of c-Myc in PANC-1 and MIA PaCa-2 cells with or without GGCT knockdown. G Cell proliferation determined by Colony assays in GGCT knockdown cells with or without overspression c-Myc. H Cell proliferation determined by CCK8 assays in GGCT knockdown cells with or without overspression c-Myc. I Assessing the progression capability of cells through Migration assays in GGCT knockdown cells with or without overspression c-Myc. * P < 0.05, ** P < 0.01, *** P < 0.001

Article Snippet: For CCK8 assay, PDAC cells (5 × 10 3 ) were cultured in 96-well plates for 0, 2, 4 days, and 10% of CCK8 reagent (TargetMol) per well were added and incubated at 37 °C for 2 h. Absorbance was measured at 450 nm using a multifunctional enzyme marker.

Techniques: Western Blot, Knockdown, Migration