Journal: Frontiers in Microbiology

Article Title: Identification of differentially expressed HERV-K(HML-2) loci in colorectal cancer

doi: 10.3389/fmicb.2023.1192900

Figure Lengend Snippet: (A–C) Comparing the HERV-K (HML-2) gag , pol , and env gene in colorectal cancer cell lines HCT116, HT29, SW480, and the human intestinal epithelial cell NCM460. Experiments were repeated three times to ensure that the results were replicable. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: The human colon mucosal epithelial cell line, NCM460, and three CRC cell lines, HCT116, HT29, and SW480, were obtained from iCell Bioscience Inc. (Shanghai, China) and Procell Life Science Technology Ltd. (Wuhan, China), respectively.

Techniques:

Journal: Journal of Gastrointestinal Oncology

Article Title: The roles and mechanisms of APOL1 in the development of colorectal cancer

doi: 10.21037/jgo-24-275

Figure Lengend Snippet: Effect of APOL1 on the biological behavior of CRC cells. (A) qRT-PCR was used to determine the APOL1 mRNA levels in HCT116, SW1116, and NCM460 cells. (B) In HCT116 cells, qRT-PCR was used to determine the interference efficacy of shAPOL1#1/2/3. (C) CCK8 assay was used to evaluate the proliferative capacity of HCT116 and SW1116 cells. (D) The count of clones in HCT116 and SW1116 cells was determined using the colony formation test. Staining method: crystal violet. Magnification: ×100. (E) Wound healing test was used to assess the migratory capacity of HCT116 and SW1116 cells (magnification: ×40). (F) The Transwell test was used to determine HCT116 and SW1116 cells’ invasion ability (magnification: ×100). Staining method: crystal violet. Each group, n=3. *, P<0.05 vs. NCM460 group; **, P<0.01; ***, P<0.001 vs . sh-NC group. sh-NC, shRNA-negative control; sh-APOL1, shRNA-apolipoprotein L1; OD, optical density; CRC, colorectal cancer; qRT-PCR, quantitative real-time polymerase chain reaction; CCK8, Cell Counting Kit 8.

Article Snippet: Shanghai iCell Bioscience Inc. (Shanghai, China) provided the human normal colon epithelial cells NCM460 (iCell-h373) and CRC cell lines [HCT116 (iCell-h071) and SW1116 cells (iCell-h201)].

Techniques: Quantitative RT-PCR, CCK-8 Assay, Clone Assay, Staining, shRNA, Negative Control, Real-time Polymerase Chain Reaction, Cell Counting

Journal: Acta Pharmaceutica Sinica. B

Article Title: Facilitation of mucosal healing by estrogen receptor β in ulcerative colitis through suppression of branched-chain amino acid transport and subsequent triggering of autophagy in colonic epithelial cells

doi: 10.1016/j.apsb.2024.11.014

Figure Lengend Snippet: ER β activation promoted focal adhesion turnover, cell migration and wound healing of colonic epithelial cells. NCM460 cells were treated with DPN (0.3, 1, and 3 μmol/L) and LIQ (3, 10, and 30 μmol/L) for 24 h. (A, B) Wound healing rate of NCM460 cells (plotting scale: 50 μm). (C, D) Proliferation rate of NCM460 cells (plotting scale: 50 μm). (E, F) Migration of NCM460 cells (plotting scale: 50 μm). (G) Confocal microscopy time-lapse sequences of NCM460 cells expressing paxillin-mCherry (white) to monitor focal adhesion dynamics (plotting scale: 1 μm). (H) Example plots of paxillin-mCherry fluorescence intensity ( y axis) over time ( x axis), and statistics of focal adhesion assembly and depolymerization time. (I, J) Expression levels of FAK and p-FAK in NCM460 cells were detected by Western blotting. The data are expressed as mean ± SEM ( n = 3). Significant differences are designated as follows: ∗ P < 0.05 and ∗∗ P < 0.01 versus the Control group.

Article Snippet: NCM460 cells transfected pmCherry- EGFP -Light chain 3 ( LC3 ) plasmid (Synbio Technologies) were seeded onto confocal dish.

Techniques: Activation Assay, Migration, Confocal Microscopy, Expressing, Fluorescence, Western Blot, Control

Journal: Acta Pharmaceutica Sinica. B

Article Title: Facilitation of mucosal healing by estrogen receptor β in ulcerative colitis through suppression of branched-chain amino acid transport and subsequent triggering of autophagy in colonic epithelial cells

doi: 10.1016/j.apsb.2024.11.014

Figure Lengend Snippet: ER β activation promoted colonic epithelial cell migration and wound healing by accelerating focal adhesion turnover. NCM460 cells were treated with DPN (1 μmol/L) for 24 h in the presence or absence of si FAK . (A) Confocal microscopy time-lapse sequences of NCM460 cells expressing paxillin-mCherry (white) to monitor focal adhesion dynamics (plotting scale: 1 μm). (B–D) Example plots of paxillin-mCherry fluorescence intensity ( y axis) over time ( x axis), and statistics of focal adhesion assembly and depolymerization time. (E) Migration of NCM460 cells (plotting scale: 50 μm). (F) Wound healing rate of NCM460 cells (plotting scale: 50 μm). The data are expressed as mean ± SEM ( n = 3). Significant differences are designated as follows: ∗∗ P < 0.01 versus the Control group; ns P > 0.05 not significant versus the si FAK group.

Article Snippet: NCM460 cells transfected pmCherry- EGFP -Light chain 3 ( LC3 ) plasmid (Synbio Technologies) were seeded onto confocal dish.

Techniques: Activation Assay, Migration, Confocal Microscopy, Expressing, Fluorescence, Control

Journal: Acta Pharmaceutica Sinica. B

Article Title: Facilitation of mucosal healing by estrogen receptor β in ulcerative colitis through suppression of branched-chain amino acid transport and subsequent triggering of autophagy in colonic epithelial cells

doi: 10.1016/j.apsb.2024.11.014

Figure Lengend Snippet: ER β activation enhanced autophagy in colonic epithelial cells and activated FAK by promoting autophagosome formation. NCM460 cells were treated with DPN (1 μmol/L) for 24 h in the presence or absence of 3-MA and CQ. (A) Volcano plot depiction of the differentially expressed genes between NCM460 cells treated with DPN and untreated NCM460 cells. (B) Scatter plots of enrichment results for the KEGG pathways and GO function enrichment for the NCM460 cells treated with DPN compared with the untreated NCM460 cells. The x -axis represents the rich factor, which calculates the proportion of differentially expressed genes to the total gene count within each KEGG or GO pathway. The color of each dot signifies the FDR value, while the dot's size corresponds to the number of genes associated with each pathway. (C) Relative changes of some representative genes in the screened autophagy signaling pathway. (D) Expression levels of LC3, ATG5, Beclin1 and SQSTM1 in NCM460 cells were detected by Western blotting. (E) Autophagosomes and autolysosomes in NCM460 cells were determined by transmission electron microscopy analysis (plotting scale: 1 μm). (F) NCM460 cells were stably transferred with pmCherry -EGFP-LC3 plasmid, and the autophagosomes (yellow fluorescence) and autolysosomes (red fluorescence) spots were observed by confocal microscopy (plotting scale: 5 μm). (G, H) Expression levels of FAK and p-FAK in NCM460 cells were detected by Western blotting. (I) The total protein of NCM460 cells was isolated and immunoprecipitated with antibody against FIP200, and the expression levels of FAK and FIP200 were analyzed by Western blotting. (J) Representative immunostaining images of NCM460 cells for FAK (red), FIP200 (green). The R represents Pearson's correlation CR coefficient value to show the signal colocalization status (plotting scale: 5 μm). The data are expressed as means ± SEM ( n = 3). Significant differences are designated as follows: ∗ P < 0.05 and ∗∗ P < 0.01 versus the Control group; ns P > 0.05 not significant versus the 3-MA group; # P < 0.05 versus the CQ group.

Article Snippet: NCM460 cells transfected pmCherry- EGFP -Light chain 3 ( LC3 ) plasmid (Synbio Technologies) were seeded onto confocal dish.

Techniques: Activation Assay, Expressing, Western Blot, Transmission Assay, Electron Microscopy, Stable Transfection, Plasmid Preparation, Fluorescence, Confocal Microscopy, Isolation, Immunoprecipitation, Immunostaining, Control

Journal: Acta Pharmaceutica Sinica. B

Article Title: Facilitation of mucosal healing by estrogen receptor β in ulcerative colitis through suppression of branched-chain amino acid transport and subsequent triggering of autophagy in colonic epithelial cells

doi: 10.1016/j.apsb.2024.11.014

Figure Lengend Snippet: ER β activation accelerated focal adhesion turnover in colonic epithelial cells by enhancing autophagy. NCM460 cells were treated with DPN (1 μmol/L) for 24 h in the presence or absence of si ATG5 . (A, B) The total protein was isolated and immunoprecipitated with antibody against FIP200, and the expression levels of FAK and FIP200 were analyzed by Western blotting. (C) Representative immunostaining images of NCM460 cells for FAK (red), FIP200 (green). The R represents Pearson's correlation CR coefficient value to show the signal colocalization status (plotting scale: 5 μm). (D) Expression levels of FAK and p-FAK in NCM460 cells were detected by Western blotting. (E) Confocal microscopy time-lapse sequences of NCM460 cells expressing paxillin-mCherry (white) to monitor focal adhesion dynamics (plotting scale: 1 μm). (F) Example plots of paxillin-mCherry fluorescence intensity ( y axis) over time ( x axis), and statistics of focal adhesion assembly and depolymerization time. The data are expressed as means ± SEM ( n = 3). Significant differences are designated as follows: ∗ P < 0.05 and ∗∗ P < 0.01 versus the Control group; ns P > 0.05 not significant versus the si ATG5 group.

Article Snippet: NCM460 cells transfected pmCherry- EGFP -Light chain 3 ( LC3 ) plasmid (Synbio Technologies) were seeded onto confocal dish.

Techniques: Activation Assay, Isolation, Immunoprecipitation, Expressing, Western Blot, Immunostaining, Confocal Microscopy, Fluorescence, Control

Journal: Acta Pharmaceutica Sinica. B

Article Title: Facilitation of mucosal healing by estrogen receptor β in ulcerative colitis through suppression of branched-chain amino acid transport and subsequent triggering of autophagy in colonic epithelial cells

doi: 10.1016/j.apsb.2024.11.014

Figure Lengend Snippet: ER β activation suppressed branched-chain amino acid transport in colonic epithelial cells by downregulating the expression of LAT1. NCM460 cells were treated with DPN (1 μmol/L) for 24 h in the presence or absence of LAT1- Plasmid. (A) Pie chart based on the main metabolic differential types between NCM460 cells treated with DPN and untreated NCM460 cells. (B) Bar chart of enrichment results for the KEGG pathways of the NCM460 cells treated with DPN compared with the untreated NCM460 cells. The x axis shows the negative log transformation of P -values. The y axis shows the pathway. (C) Relative changes of some representative metabolites in the screened amino acids transport pathway. (D) The levels of leucine, isoleucine and valine in NCM460 cells were detected by ELISA kits. (E) Expression level of LAT1 in NCM460 cells was detected by Western blotting. (F) The DNA sequence of LAT1 , marked with five silencers. (G) Schematic diagram of the Cut & Tag experimental, sourced from Vazyme Biotech Co., Ltd. Expression of LAT1 silencers binding with ER β was detected by Cut & Tag-qPCR. (H) Expression levels of p-mTORC1 and mTORC1 in NCM460 cells were detected by Western blotting. The data are expressed as mean ± SEM ( n = 3). Significant differences are designated as follows: ∗ P < 0.05 and ∗∗ P < 0.01 versus the Control group.

Article Snippet: NCM460 cells transfected pmCherry- EGFP -Light chain 3 ( LC3 ) plasmid (Synbio Technologies) were seeded onto confocal dish.

Techniques: Activation Assay, Expressing, Plasmid Preparation, Transformation Assay, Enzyme-linked Immunosorbent Assay, Western Blot, Sequencing, Binding Assay, Control

Journal: Acta Pharmaceutica Sinica. B

Article Title: Facilitation of mucosal healing by estrogen receptor β in ulcerative colitis through suppression of branched-chain amino acid transport and subsequent triggering of autophagy in colonic epithelial cells

doi: 10.1016/j.apsb.2024.11.014

Figure Lengend Snippet: ER β activation promoted autophagy in colonic epithelial cells by suppressing branched-chain amino acid transport. NCM460 cells were treated with DPN (1 μmol/L) for 24 h in the presence or absence of LAT1 -Plasmid. (A) The levels of leucine, isoleucine and valine in NCM460 cells were detected by ELISA kits. (B) Expression levels of LC3, ATG5, Beclin1 and SQSTM1 in NCM460 cells were detected by Western blotting. (C) Autophagosomes and autolysosomes in NCM460 cells were determined by transmission electron microscopy analysis (plotting scale: 1 μm). (D) NCM460 cells were stably transferred with pmCherry -EGFP - LC3 plasmid, and the autophagosomes (yellow fluorescence) and autolysosomes (red fluorescence) spots were observed by confocal microscopy (plotting scale: 5 μm). The data are expressed as mean ± SEM ( n = 3). Significant differences are designated as follows: ∗ P < 0.05 and ∗∗ P < 0.01 versus the Control group; ns P > 0.05 not significant versus the LAT1- Plasmid group.

Article Snippet: NCM460 cells transfected pmCherry- EGFP -Light chain 3 ( LC3 ) plasmid (Synbio Technologies) were seeded onto confocal dish.

Techniques: Activation Assay, Plasmid Preparation, Enzyme-linked Immunosorbent Assay, Expressing, Western Blot, Transmission Assay, Electron Microscopy, Stable Transfection, Fluorescence, Confocal Microscopy, Control

Journal: Scientific Reports

Article Title: Toxic metabolite profiling of Inocybe virosa

doi: 10.1038/s41598-020-70196-7

Figure Lengend Snippet: ( A ) The dose–response effect of IV extract on NCM460 cells; IC 50 value—0.63 mg/mL, ( B ) the effect of IV extract on the morphology of NCM460 cells under phase contrast microscope (20 ×).

Article Snippet: The immortalized cell lines, NCM460 and Chang liver cell line were both obtained from National Centre for Cell Sciences (NCCS), Pune, India and grown at 37 °C in a humidified atmosphere of 5% CO 2 using MEM (Minimum Essential Media), supplemented with 10% FBS and antibiotic solution.

Techniques: Microscopy