Journal: Innate immunity

Article Title: LncRNA GAS5 enhanced the killing effect of NK cell on liver cancer through regulating miR-544/RUNX3.

doi: 10.1177/1753425919827632

Figure Lengend Snippet: Figure 3. lncRNA GAS5 knockdown inhibited the killing effect of NK cells. NK92 cells were activated by IL-2 (100 U/ml) and transfected with si-control or si-GAS5. (a) lncRNA GAS5 was down-regulated in the si-GAS5 group compared with the si-control group. (b) IFN-c secretion in the supernatant was decreased in the si-GAS5 group compared with the si-control group. (c) After transfection with si-control or si-GAS5, NK cells were co-cultured with HepG2 cells, and cytotoxicity was detected by the CytoTox 96 non-radioactive cytotoxicity assay. Cytotoxicity was decreased in the si-GAS5 group compared with the si-control group. After co- culturing NK92 cells and Huh7 cells, cytotoxicity was decreased in the si-GAS5 group compared with the si-control group. (d) Lysosome marker CD107a of NK cells was detected by flow cytometry. The percentage of CD107aþ of NK cells was decreased in the si-GAS5 group compared with the si-control group. (e) Annexin V-PI double staining assay was used to detect the apoptosis of HepG2 and Huh7 cells. HepG2 cells were divided into si-control and si-GAS5 groups, and the apoptosis rate in si-GAS5 group was reduced compared with si-control group (41% vs 21.49%). Huh7 cells were divided into si-control and si-GAS5 groups, and the apoptosis rate in si-GAS5 group was reduced compared with si-control group (36.45% vs 13.25%). *P < 0.05, compared with si-control.

Article Snippet: Co-culture of NK92 cells and HepG2 cells or Huh7 cells Human hepatic cancer cell line HepG2 or Huh7 was purchased from ATCC, and 1.5 105 cells/ml diluted with DMEM medium (Gibco) supplemented with 10% FBS were seeded in 96-well plates at 100 ll/well.

Techniques: Knockdown, Transfection, Control, Cell Culture, Cytotoxicity Assay, Marker, Flow Cytometry, Double Staining

Journal: Innate immunity

Article Title: LncRNA GAS5 enhanced the killing effect of NK cell on liver cancer through regulating miR-544/RUNX3.

doi: 10.1177/1753425919827632

Figure Lengend Snippet: Figure 5. lncRNA GAS5 enhanced the killing effect of NK cells through miR-544/RUNX3. NK92 cells were activated by IL-2 (100 U/ml), transfected with lenti-GAS5 or miR-544 mimic, and divided into lenti-NC, lenti-GAS5, lenti-GAS5 þ pre-NC, lenti- GAS5 þ miR-544 mimic groups. (a) GAS5 expression was up-regulated in the lenti-GAS5 group compared with the lenti-NC group, and an miR-544 mimic did not change the GAS5 level. (b) Protein level of RUNX3 was up-regulated in the lenti-GAS5 group compared with the lenti-NC group, and an miR-544 mimic reversed the promotion effect of lenti-GAS5. (c) IFN-c secretion in the supernatant was increased in the lenti-GAS5 group, and miR-544 mimic reversed the promotion effect of lenti-GAS5. (d) After transfection, NK cells were co-cultured with HepG2 cells. Cytotoxicity was enhanced in the lenti-GAS5 group, and an miR-544 mimic reversed the promotion effect of lenti-GAS5. (e) The percentage of CD107aþ of NK cells was increased in the lenti-GAS5 group, and an miR-544 mimic reversed the promotion effect of lenti-GAS5. (f) Annexin V-PI double staining assay showed that apoptosis rate of HepG2 cells in lenti-NC, lenti-GAS5, lenti-GAS5 þ pre-NC, or lenti-GAS5 þ miR-544 mimic was 41.83%, 53.61%, 58.84%, and 44.58%, respec- tively. *P < 0.05, compared with lenti-NC; #P < 0.05, compared with lenti-GAS5 þ pre-NC.

Article Snippet: Co-culture of NK92 cells and HepG2 cells or Huh7 cells Human hepatic cancer cell line HepG2 or Huh7 was purchased from ATCC, and 1.5 105 cells/ml diluted with DMEM medium (Gibco) supplemented with 10% FBS were seeded in 96-well plates at 100 ll/well.

Techniques: Transfection, Expressing, Cell Culture, Double Staining

Journal: Innate immunity

Article Title: LncRNA GAS5 enhanced the killing effect of NK cell on liver cancer through regulating miR-544/RUNX3.

doi: 10.1177/1753425919827632

Figure Lengend Snippet: Figure 6. Xenograft in nude mice. HepG2 cells (6 106 cells) were injected subcutaneously into BALB/c nude mice. NK cells transfected with lenti-GAS5 or lenti-NC were injected intravenously twice at 2 h after HepG2 implantation and at d 7, and nude mice were divided into lenti-GAS5 or lenti-NC groups, with six mice in each group. (a) Tumor volume was measured every 5 d, and the nude mice were sacrificed at d 20. (b) lncRNA GAS5 was up-regulated in the lenti-GAS5 group. C. miR-544 was down-regulated in the lenti-GAS5 group. (d) RUNX3 was up-regulated in the lenti-GAS5 group. *P < 0.05, compared with lenti-NC.

Article Snippet: Co-culture of NK92 cells and HepG2 cells or Huh7 cells Human hepatic cancer cell line HepG2 or Huh7 was purchased from ATCC, and 1.5 105 cells/ml diluted with DMEM medium (Gibco) supplemented with 10% FBS were seeded in 96-well plates at 100 ll/well.

Techniques: Injection, Transfection

Journal: Nature Microbiology

Article Title: Cryptosporidium PI(4)K inhibitor EDI048 is a gut-restricted parasiticidal agent to treat paediatric enteric cryptosporidiosis

doi: 10.1038/s41564-024-01810-x

Figure Lengend Snippet: Anti- Cryptosporidium activity, in vitro and in vivo parameters of soft drug and other compounds

Article Snippet: Cytotoxicity against HepG2 ATCC CRL-10741 was determined as previously described .

Techniques: Activity Assay, In Vitro, In Vivo, Solubility, Permeability, Clinical Proteomics

Journal: Journal of molecular structure

Article Title: Synthesis, biological evaluation, and modeling studies of 1,3-disubstituted cyclobutane-containing analogs of combretastatin A4

doi: 10.1016/j.molstruc.2020.128025

Figure Lengend Snippet: IC 50 values (mean; n=4) of reference and studied compounds to HepG2 and SK-N-DZ cell lines.

Article Snippet: Cytotoxity assay Human hepatocellular carcinoma cell line HepG2 and neuroblastoma cell line SK-N-DZ were provided by Promega, USA.

Techniques:

Journal: Journal of molecular structure

Article Title: Synthesis, biological evaluation, and modeling studies of 1,3-disubstituted cyclobutane-containing analogs of combretastatin A4

doi: 10.1016/j.molstruc.2020.128025

Figure Lengend Snippet: Dose-response curves for the compounds evaluated in resazurin-based cell viability assay against HepG2 cells (mean ± SD, n = 4).

Article Snippet: Cytotoxity assay Human hepatocellular carcinoma cell line HepG2 and neuroblastoma cell line SK-N-DZ were provided by Promega, USA.

Techniques: Viability Assay

Journal: Laboratory Investigation; a Journal of Technical Methods and Pathology

Article Title: Bile acid-induced tissue factor activity in hepatocytes correlates with activation of farnesoid X receptor

doi: 10.1038/s41374-021-00628-z

Figure Lengend Snippet: TF activity was assessed via the generation of coagulation factor Xa (FXa). A HepG2 cells were incubated with CDCA, GCDCA, and UCDA (100–600 µM) for 24 h. B HepG2 cells were incubated with GW4064 (1–40 µM) for 24 h. Dimethyl sulfoxide was used as the vehicle. Data are expressed as mean ± SD (N = 6). C MTT assay showed the viability of the cells at the concentrations used in the TF activity assay. HepG2 cells were incubated with CDCA, GCDCA, and UDCA (125–4000 µM) for 24 h. D HepG2 cells were incubated with GW4064 (1–100 µM) for 24 h. Data are presented as % viability and expressed as mean ± SD (N = 6). E/F Cytotoxicity assay showed no observable LDH release with CDCA, GCDCA, UDCA, and GW4064 after 24 h. Data are presented as % cytotoxicity and expressed as mean ± SD (N = 6). PC positive control consisting of lysed cells. *p < 0.05 for a difference from a vehicle; **p < 0.001 for a difference from a vehicle.

Article Snippet: Cell culture HepG2 cells (source: cell culture core facility, Zentrum für Medizinische Forschung, Medizinische Universität Graz) ( Center for Medical Research, Medical University of Graz; cells originally derived from CLS Cell Lines Service, Eppelheim, Germany, and Short Tandem Repeat-verified before usage) were cultured in Dulbecco’s modified Eagle’s medium (DMEM, GIBCO, Thermo Fisher Scientific, Waltham, MA), supplemented with 10% fetal bovine serum (GIBCO) and 1% penicillin-streptomycin (GIBCO) at 37 °C and 5% CO 2 .

Techniques: Activity Assay, Coagulation, Incubation, MTT Assay, Cytotoxicity Assay, Positive Control

Journal: Laboratory Investigation; a Journal of Technical Methods and Pathology

Article Title: Bile acid-induced tissue factor activity in hepatocytes correlates with activation of farnesoid X receptor

doi: 10.1038/s41374-021-00628-z

Figure Lengend Snippet: TF activity was assessed via generation of coagulation factor Xa (FXa) after exposure of HepG2 cells to (A) CDCA (100–600 µM) or (C) GW4064 (10–60 µM) for 24 h with and without the addition of the FXR inhibitor DY 268 (N = 3). B/D MTT assay showed unimpaired viability. **p < 0.001 for the difference from the vehicle.

Article Snippet: Cell culture HepG2 cells (source: cell culture core facility, Zentrum für Medizinische Forschung, Medizinische Universität Graz) ( Center for Medical Research, Medical University of Graz; cells originally derived from CLS Cell Lines Service, Eppelheim, Germany, and Short Tandem Repeat-verified before usage) were cultured in Dulbecco’s modified Eagle’s medium (DMEM, GIBCO, Thermo Fisher Scientific, Waltham, MA), supplemented with 10% fetal bovine serum (GIBCO) and 1% penicillin-streptomycin (GIBCO) at 37 °C and 5% CO 2 .

Techniques: Activity Assay, Coagulation, MTT Assay

Journal: Laboratory Investigation; a Journal of Technical Methods and Pathology

Article Title: Bile acid-induced tissue factor activity in hepatocytes correlates with activation of farnesoid X receptor

doi: 10.1038/s41374-021-00628-z

Figure Lengend Snippet: The ability of a sample to generate thrombin was evaluated with respect to the time preceding the thrombin burst (lag time) and the total amount of thrombin activity (endogenous thrombin potential), which was assessed as the area under the curve. HepG2 cells were incubated with 100, 200, 400, or 600 μM of (A) CDCA, (B) GCDCA, (C) UDCA, or (D) 1, 5, 10, or 40 μM of GW4064. Dimethyl sulfoxide was used as the vehicle. Measurements were carried out after the addition of corn trypsin inhibitor to block contact activation of coagulation. E Lag time reduction/prolongation in %, induced by various bile acids in different concentrations. Data are expressed as mean ± SD (N = 9). **p < 0.01 for the difference from the vehicle.

Article Snippet: Cell culture HepG2 cells (source: cell culture core facility, Zentrum für Medizinische Forschung, Medizinische Universität Graz) ( Center for Medical Research, Medical University of Graz; cells originally derived from CLS Cell Lines Service, Eppelheim, Germany, and Short Tandem Repeat-verified before usage) were cultured in Dulbecco’s modified Eagle’s medium (DMEM, GIBCO, Thermo Fisher Scientific, Waltham, MA), supplemented with 10% fetal bovine serum (GIBCO) and 1% penicillin-streptomycin (GIBCO) at 37 °C and 5% CO 2 .

Techniques: Activity Assay, Incubation, Blocking Assay, Activation Assay, Coagulation

Journal: Laboratory Investigation; a Journal of Technical Methods and Pathology

Article Title: Bile acid-induced tissue factor activity in hepatocytes correlates with activation of farnesoid X receptor

doi: 10.1038/s41374-021-00628-z

Figure Lengend Snippet: A–D Representative Western blots of TF after incubation of HepG2 cells with CDCA, GCDCA, UDCA (100–600 µM), or GW4064 (10–60 µM). Dimethyl sulfoxide was used as the vehicle. GAPDH was used as a control. E Densitometric analysis with normalization to GAPDH from three independent experimental replicates.

Article Snippet: Cell culture HepG2 cells (source: cell culture core facility, Zentrum für Medizinische Forschung, Medizinische Universität Graz) ( Center for Medical Research, Medical University of Graz; cells originally derived from CLS Cell Lines Service, Eppelheim, Germany, and Short Tandem Repeat-verified before usage) were cultured in Dulbecco’s modified Eagle’s medium (DMEM, GIBCO, Thermo Fisher Scientific, Waltham, MA), supplemented with 10% fetal bovine serum (GIBCO) and 1% penicillin-streptomycin (GIBCO) at 37 °C and 5% CO 2 .

Techniques: Western Blot, Incubation, Control

Journal: Laboratory Investigation; a Journal of Technical Methods and Pathology

Article Title: Bile acid-induced tissue factor activity in hepatocytes correlates with activation of farnesoid X receptor

doi: 10.1038/s41374-021-00628-z

Figure Lengend Snippet: Caspase 3 activity was determined with a colorimetric assay based on p-nitroaniline (pNA) release from a peptide substrate (Ac-DEVD-pNA). HepG2 cells were incubated with CDCA, GCDCA, and UDCA (100–600 μM), GW4064 (1–60 µM), or staurosporine (2–4 µM) for 24 h. Dimethyl sulfoxide was used as a vehicle. Data are expressed as mean ± SD (N = 6). **p < 0.01 for a difference from the vehicle.

Article Snippet: Cell culture HepG2 cells (source: cell culture core facility, Zentrum für Medizinische Forschung, Medizinische Universität Graz) ( Center for Medical Research, Medical University of Graz; cells originally derived from CLS Cell Lines Service, Eppelheim, Germany, and Short Tandem Repeat-verified before usage) were cultured in Dulbecco’s modified Eagle’s medium (DMEM, GIBCO, Thermo Fisher Scientific, Waltham, MA), supplemented with 10% fetal bovine serum (GIBCO) and 1% penicillin-streptomycin (GIBCO) at 37 °C and 5% CO 2 .

Techniques: Activity Assay, Colorimetric Assay, Incubation

Journal: Laboratory Investigation; a Journal of Technical Methods and Pathology

Article Title: Bile acid-induced tissue factor activity in hepatocytes correlates with activation of farnesoid X receptor

doi: 10.1038/s41374-021-00628-z

Figure Lengend Snippet: HepG2 cells were incubated with 200 μM of chenodeoxycholic acid (CDCA) and with 20 μM of GW4064 for 12 h or 24 h. SHP, OST-α, OST-ß, and tissue factor (TF) were used as primers, with SHP, OST-α, and OST-ß as known FXR targets. Data were normalized using the expression of β-actin and relative gene expression was assessed using the ΔΔCq method. ΔΔCq-values were normalized to dimethyl sulfoxide. Data are expressed as mean fold change (2−ΔΔCq) (N = 2).

Article Snippet: Cell culture HepG2 cells (source: cell culture core facility, Zentrum für Medizinische Forschung, Medizinische Universität Graz) ( Center for Medical Research, Medical University of Graz; cells originally derived from CLS Cell Lines Service, Eppelheim, Germany, and Short Tandem Repeat-verified before usage) were cultured in Dulbecco’s modified Eagle’s medium (DMEM, GIBCO, Thermo Fisher Scientific, Waltham, MA), supplemented with 10% fetal bovine serum (GIBCO) and 1% penicillin-streptomycin (GIBCO) at 37 °C and 5% CO 2 .

Techniques: Incubation, Expressing, Gene Expression

Journal: Laboratory Investigation; a Journal of Technical Methods and Pathology

Article Title: Bile acid-induced tissue factor activity in hepatocytes correlates with activation of farnesoid X receptor

doi: 10.1038/s41374-021-00628-z

Figure Lengend Snippet: Parameters of thrombin generation curves after incubation of HepG2 cells with various bile acids or GW4064 ( N = 6).

Article Snippet: Cell culture HepG2 cells (source: cell culture core facility, Zentrum für Medizinische Forschung, Medizinische Universität Graz) ( Center for Medical Research, Medical University of Graz; cells originally derived from CLS Cell Lines Service, Eppelheim, Germany, and Short Tandem Repeat-verified before usage) were cultured in Dulbecco’s modified Eagle’s medium (DMEM, GIBCO, Thermo Fisher Scientific, Waltham, MA), supplemented with 10% fetal bovine serum (GIBCO) and 1% penicillin-streptomycin (GIBCO) at 37 °C and 5% CO 2 .

Techniques: Incubation