Journal: Journal of Cancer

Article Title: Tranexamic acid reduces endometrial cancer effects through the production of angiostatin

doi: 10.7150/jca.68169

Figure Lengend Snippet: Effects of TA treatment on plasminogen, plasmin and tPA levels. Thirty weeks after the start of the study, by western blotting, the expression of plasminogen (A) , plasmin (B) and tPA (C) in uterus on MNU- and estradiol-induced endometrial cancer model mice were measured. By ELISA kit, the levels of plasminogen (D) , plasmin (E) and tPA (F) in plasma on MNU- and estradiol-induced endometrial cancer model mice were measured. Values are expressed as the mean ± standard deviation (SD) derived from 10 animals. * p < 0.05. TA: tranexamic acid, tPA: tissue plasminogen activator, MNU: N -methyl- N -nitrosourea.

Article Snippet: The plasma levels of carbohydrate antigen 125 (CA125), interleukin (IL)-6, tumor necrosis factor (TNF-α), plasminogen, plasmin, and tPA were determined using commercial ELISA kits (CA125: Bioassay Technology Laboratory, Shanghai, China; Il-6: Proteintech, Rosemont, IL, USA; TNF-α: R&D Systems, Minneapolis, MN, USA; plasminogen and plasmin: LSBio, Seattle, WA, USA; tPA: Abcam, Cambridge, UK) according to the respective manufacturers' instructions.

Techniques: Western Blot, Expressing, Enzyme-linked Immunosorbent Assay, Clinical Proteomics, Standard Deviation, Derivative Assay

Journal: Journal of Translational Medicine

Article Title: Hypoxic BMSCs accelerate femur fracture healing via the METTL3/SLC2A3 m6A-glycolysis axis

doi: 10.1186/s12967-025-07510-2

Figure Lengend Snippet: Hypoxia induces higher osteogenic capacity in BMSCs . ( A ) Flow cytometry assesses the expression levels of BMSCs markers (CD29 and CD44) and another cell marker (CD45). ( B ) CCK-8 assay was conducted to assess the cell viability of BMSCs after stimulation with different concentrations of CoCl₂ for 72 h. * P < 0.05, ** P < 0.01, *** P < 0.001, one-way ANOVA. ( C ) Alizarin Red staining detects the osteogenic capacity. ( D ) Immunofluorescence assay showed the expression level of HIF1α. ( E ) ELISA analysis of ALP levels in Ctrl and Hypoxia groups. ( F ) Runx2, OCN, and Osx biomarkers were assessed using WB. *** P < 0.001, Student’s t -test

Article Snippet: BMSC identity was confirmed by flow cytometry (NovoCyte, Agilent, USA) using antibodies against CD29 (No. 130-119-165), CD44 (No. 130-107-802), and CD45 (No. 130-128-256; 1:100 diluted, Miltenyi Biotech, Germany).

Techniques: Flow Cytometry, Expressing, Marker, CCK-8 Assay, Staining, Immunofluorescence, Enzyme-linked Immunosorbent Assay

Journal: bioRxiv

Article Title: Mycobacterium tuberculosis SecA2-dependent activation of host Rig-I/MAVs signaling is not conserved in Mycobacterium marinum

doi: 10.1101/2023.01.27.525853

Figure Lengend Snippet: M. marinum strains were grown to exponential phase in 7H9 supplemented with 10% OADC and 0.2% tyloxapol before being subcultured into fresh media at an optical density (OD 600 ) of 0.05. Bacterial growth was monitored by measuring the OD 600 every 24-hours for 7 days.

Article Snippet: Mycobacterium marinum M bacterial strains were grown on Middlebrook’s 7H11 (Sigma-Aldrich) agar plates supplemented with 10% OADC [oleic acid (Fischer Scientific), dextrose (Sigma and VWR), albumin (Sigma Aldrich), and catalase(Fischer)] and 0.5% glycerol (VWR, Radnor, PA) at 30°C or in Middlebrook’s 7H9 media (Sigma-Aldrich, St. Louis, MO) supplemented with 10% OADC and 0.2% tyloxapol (Chem-Impex Int’l Inc., Wood Dale, IL) at 30°C.

Techniques:

Journal: bioRxiv

Article Title: Mycobacterium tuberculosis SecA2-dependent activation of host Rig-I/MAVs signaling is not conserved in Mycobacterium marinum

doi: 10.1101/2023.01.27.525853

Figure Lengend Snippet: M. marinum strains were grown to exponential phase in 7H9 supplemented with 10% OADC and 0.2% tyloxapol before being subcultured into fresh media at an optical density (OD 600 ) of 0.05. Bacterial growth was monitored by measuring the OD 600 every 24-hours for 7 days and plotted on a log transformed scale (A). The same strains were examined for growth in sodium dodecyl-sulfate (SDS) by subculturing exponential phase bacteria into 7H9 supplemented with 10% OADC and 0.2% tyloxapol with and without 0.2% SDS at an OD600 of 0.8. Twenty-four hours later, these cultures were serially diluted and plated onto 7H11 agar plates supplemented with 10% OADC in technical triplicate. Images are representative of three biological replicates each plated in technical triplicate (B). Colony forming units were quantified from agars plates following incubation at 32°C with 5% CO 2 for one week (C). Statistical significance was determined using a one-way ANOVA followed by a Dunnett’s test for multiple comparison relative to WT, with p-values ≤ 0.05 considered significant (C only).

Article Snippet: Mycobacterium marinum M bacterial strains were grown on Middlebrook’s 7H11 (Sigma-Aldrich) agar plates supplemented with 10% OADC [oleic acid (Fischer Scientific), dextrose (Sigma and VWR), albumin (Sigma Aldrich), and catalase(Fischer)] and 0.5% glycerol (VWR, Radnor, PA) at 30°C or in Middlebrook’s 7H9 media (Sigma-Aldrich, St. Louis, MO) supplemented with 10% OADC and 0.2% tyloxapol (Chem-Impex Int’l Inc., Wood Dale, IL) at 30°C.

Techniques: Transformation Assay, Subculturing Assay, Bacteria, Incubation, Comparison

Journal: bioRxiv

Article Title: Mycobacterium tuberculosis SecA2-dependent activation of host Rig-I/MAVs signaling is not conserved in Mycobacterium marinum

doi: 10.1101/2023.01.27.525853

Figure Lengend Snippet: M. marinum strains were grown to exponential phase in 7H9 supplemented with 10% OADC and 0.2% tyloxapol and used to infect triplicate wells of BMDMs, at an MOI of 0.2, for 2 hours. At 2, 48, and 96 hours post infection (hpi), macrophage monolayers were lysed and plated onto 7H11 agar supplemented with 10% OADC in technical triplicate. One week after plating, colony forming units were enumerated for each timepoint (A). Bacterial numbers present at 2hpi were plotted separately (B) to better visualize differences in bacterial counts. Uninfected wells were included as a control for across well contamination (not shown, no contamination observed) and the Δ esxBA strain as a control for attenuated growth. Statistical significance was determined using a Kruskal-Wallis test followed by a Wilcoxon Rank Sum test for pairwise comparison relative to WT, with p-values ≤ 0.05 considered significant (*** p<0.001).

Article Snippet: Mycobacterium marinum M bacterial strains were grown on Middlebrook’s 7H11 (Sigma-Aldrich) agar plates supplemented with 10% OADC [oleic acid (Fischer Scientific), dextrose (Sigma and VWR), albumin (Sigma Aldrich), and catalase(Fischer)] and 0.5% glycerol (VWR, Radnor, PA) at 30°C or in Middlebrook’s 7H9 media (Sigma-Aldrich, St. Louis, MO) supplemented with 10% OADC and 0.2% tyloxapol (Chem-Impex Int’l Inc., Wood Dale, IL) at 30°C.

Techniques: Infection, Control, Comparison

Journal: International Journal of Molecular Sciences

Article Title: Reversible Growth-Arrest of a Spontaneously-Derived Human MSC-Like Cell Line

doi: 10.3390/ijms21134752

Figure Lengend Snippet: Flow cytometry analysis of various expression markers was performed in 12 different human MSC populations, six derived from umbilical cord (UC) in passage (P) 0 to P3 (blue background), three from bone marrow (BM) in P2 to P4 (red background), three from placenta (PL) (green background) in P1 to P2, and compared to MSC544 in P22 (yellow background). Proteins at low to undetectable levels are displayed by a khaki-colored background.

Article Snippet: After blocking with 2 % Fetal calf serum (FCS) for 15 min at room temperature and subsequent washing in PBS the cells were stained with 7-AAD (7-aminoactinomycin D) viability staining solution (BioLegend via Biozol GmbH, Eching, Germany) and the following monoclonal anti-human APC- (allophycocyanin), PB- (pacific blue), PE- (phycoerythrin) or FITC- (fluorescein isothiocyanate) labeled antibodies: CD73-PE and CD73-PB (clone AD2, BD Bioscience GmbH, Heidelberg, Germany); CD90-PE and CD90-APC (clone 5E10, BioLegend via Biozol GmbH); CD105-PE (clone 43A3, BioLegend via Biozol GmbH); rat CD146-PE (clone ME-09F1, Miltenyi Biotech GmbH, Bergisch Gladbach, Germany); CD166-PE (clone REA442, Miltenyi Biotech GmbH); CD326-PE (clone HEA-125, Miltenyi Biotech GmbH); CD31-FITC (clone WM59, Dako, Agilent, Santa Clara, CA, USA); CD13-PE (clone WM15, BioLegend via Biozol GmbH); CD29PE (clone MAR4, BD Bioscience GmbH); CD44-FITC (clone G44-26, BD Bioscience GmbH); CA125 (clone X75, Abcam, Cambridge, UK); CD54-PE (clone HCD54, BioLegend via Biozol GmbH); CD106-PE (clone STA, BioLegend via Biozol GmbH); CD24-FITC (clone ML5, BD Bioscience GmbH); CD14-PE (clone TÜK4, Miltenyi Biotech GmbH); CD45-PE (clone HI30, BioLegend via Biozol GmbH); CD133-PE (clone293C3, Miltenyi Biotech GmbH); CD271-PE (clone ME20.4-1.H4, Miltenyi Biotech GmbH); mesothelin-PE (clone REA1057, Miltenyi Biotech GmbH).

Techniques: Flow Cytometry, Expressing, Derivative Assay, Labeling