Journal:

Article Title: Mycobacterium bovis BCG-Infected Mice Are More Susceptible to Staphylococcal Enterotoxin B-Mediated Toxic Shock than Uninfected Mice despite Reduced In Vitro Splenocyte Responses to Superantigens †

doi: 10.1128/IAI.70.8.4148-4157.2002

Figure Lengend Snippet: BCG infectious dose correlates with increased expression of the T-cell activation markers CD44, IL-2Rα, and IL-2Rβ. Unfractionated splenocytes (5 × 105 cells/tube) from each type of spleen were stained with FITC-labeled anti-mouse CD44, anti-mouse CD25/IL-2Rα, or anti-mouse CD122/IL-2Rβ and PE-labeled anti-mouse CD4 or anti-mouse CD8α. The percentages of CD4+ CD44hi, IL-2Rαhi, or IL-2Rβhi CD4+ (white bars) or CD8+ cells (black bars) in the spleen were determined by flow cytometry. Means ± standard errors of the means of three experiments are shown. ∗, P < 0.05; ∗∗, P < 0.01. Purified CD4+ T cells from the spleens of BCG HD-infected mice (day 16) were stained with FITC anti-mouse CD44 and were sorted in an EPICS Elite ESP (Beckman Coulter) into CD44low (naïve) and CD44hi (memory/effector) populations. Sorted cells were cocultured at 105 cells/well either with bone marrow-derived dendritic cells (5 × 104 cells/well) in the presence or absence of SEB (10 μg/ml) or with M12 B-lymphoma cells (5 × 104 cells/well) in the presence or absence of BCG-Ag (10 μg/ml). IFN-γ levels were determined as described in Materials and Methods. Mean ± standard deviation of triplicate wells is shown.

Article Snippet: For flow cytometry analysis, splenocytes (5 × 10 5 ) were stained with all or some of the following MAbs (all at 1:100 dilutions in R10A): fluorescein isothiocyanate (FITC)-conjugated mouse anti-mouse Vβ8 TCR (F23.1), FITC-conjugated rat anti-mouse CD25 (IL-2 Rα; 7D4), FITC-conjugated rat anti-mouse CD122 (IL-2 Rβ; TM-β1), FITC-conjugated rat anti-mouse I-A d /I-E d (2G9) (all from PharMingen), or FITC-conjugated rat anti-mouse CD44 (IM7.8.1; Cedarlane Laboratories, Hornby, Canada).

Techniques: Expressing, Activation Assay, Staining, Labeling, Flow Cytometry, Purification, Infection, Derivative Assay, Standard Deviation

Journal: Discover Oncology

Article Title: Preclinical efficacy of CBR-5884 against epithelial ovarian cancer cells by targeting the serine synthesis pathway

doi: 10.1007/s12672-024-01013-0

Figure Lengend Snippet: CBR-5884-impaired stemness of EOC cells and enhanced chemotherapy sensitivity. A SKOV3 and ID8 cells were treated with CBR-5884 (50 μM) or dimethyl sulfoxide (DMSO; control) for 5 days, and tumorsphere formation ability was accessed by sphere formation assay. Representative images (left) and quantification data (right) are shown. B Cell cytometry was performed to detect CD44 expression in SKOV3 and ID8 cells after treatment with 30 μM CBR-5884 for 48 h. Representative images (left) and quantification data (right) are shown. C Quantitative polymerase chain reaction assay was conducted to measure the mRNA expression levels of stem-related genes in SKOV3 cells, including ALDH1A1 , NANOG , SOX2 , EPCAM , and POU5F1 after treatment with 30 μM CBR-5884 for 48 h. D SKOV3 and ID8 cells were treated with DMSO, carboplatin (20 μM), CBR-5884 (30 μM), and carboplatin (20 μM) plus CBR-5884 (30 μM) for 72 h. Cell viability was measured using CCK-8 assay

Article Snippet: To detect CD44-positive cells, a FITC anti-mouse CD44 (IM7; 1:200 dilution, Cat. No. FITC-65117; Proteintech, USA) antibody was used.

Techniques: Control, Tube Formation Assay, Cytometry, Expressing, Real-time Polymerase Chain Reaction, CCK-8 Assay