Journal: Neuro-Oncology

Article Title: Dissecting inherent intratumor heterogeneity in patient-derived glioblastoma culture models

doi: 10.1093/neuonc/now253

Figure Lengend Snippet: Floating cells display enhanced mesenchymal properties. (A) MGG6 NS and FC mRNA were analyzed by qRT-PCR for different mesenchymal markers including CD44, vimentin, YKL-40, N-cad, and MMP2. (B) Cell lysates from different cultures were analyzed by western blotting for pSTAT3, STAT3, CD44, C/EBPβ, and β-actin. (C) FACS analysis showing proportion of CD44 positive cells in FC and their corresponding NS from different GSC cultures. (D) MGG29 NS were fractionated into CD44high and CD44low subpopulation by FACS and predominant cells were established and labeled with GFP and mCherry, respectively. Left: fluorescence analysis of a mixture of these 2 subpopulations cultured in serum; middle: FC collected and analyzed for both GFP and mCherry; right: flow cytometry of CD44high cells in both GFP and mCherry populations. (E) NS and FC were differentiated using StemPro Chondrogenesis Differentiation Kit and stained with fast green and safranin O (positive, #; negative, +). (F) NS and FC were cultured as a monolayer and their ability to invade/migrate was evaluated using scratch healing assay. Bright field micrographs are showing for different groups. Scale bar, 50 μm.

Article Snippet: CD44 expression was measured in 500000 cells dissociated with 2 mM EDTA and incubated in 80 μL PBS/0.1% bovine serum albumin at 4°C for 30 min in the dark with mouse monoclonal anti-human CD44 antibody conjugated to fluorescein isothiocyanate (FITC, 1:10; Miltenyi Biotec) or respective mouse immunoglobulin G1 control in the presence of FcR blocking reagent (1:5).

Techniques: Quantitative RT-PCR, Western Blot, Labeling, Fluorescence, Cell Culture, Flow Cytometry, Staining

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: Fig. 5 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 Quanti- tative polymerase chain reaction assay was conducted to measure the mRNA expression levels of stem-related genes in SKOV3 cells, includ- ing 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, Polymerase Chain Reaction, CCK-8 Assay