ca125  (Santa Cruz Biotechnology)

Journal: Biomaterials science

Article Title: Self-targeted nanosystem for enhanced chemodynamic cancer therapy.

doi: 10.1039/d4bm01683a

Figure Lengend Snippet: Fig. 6 (a and b) Western blot analysis of oxidative genes. (a) A2780 and (b) ID8 cells were incubated with ZnNP, ZnCl2, SNP (25 μg ml−1), and CisPt (3 µg ml−1; 10 µM) for 24 h. (c–f) Cytotoxicity of ZnNP, ZnCl2, SNP and CisPt on healthy liver and PDTOs obtained from HGSOC patients (Pat A–C). Error bars represent the standard error of the mean (SEM). Groups were considered statistically significant if p < 0.05 (*), p < 0.01 (**) or non-signifi- cant (ns). (g) Hematoxylin & eosin staining and immunohistochemistry (PAT A–C). CA-125 (cancer antigen 125), WT1 (Wilms’ tumor 1) and PAX8 (paired box gene 8) are markers of ovarian cancer (all images 40×, bar = 50 µm).

Article Snippet: The following antibodies were used to characterize PDTO and parental tumours: PAX8 (catalog no.: 10336-1-AP, dilution 1 : 400, ProteinTech Group, Planegg-Martinsried, Germany), CA125 (catalog no.: sc-52095, dilution 1 : 100, Santa Cruz Biotechnology, Dallas, TX, USA) and WT1 (catalog no.: ab89901, dilution 1 : 300, Abcam, Cambridge, UK).

Techniques: Western Blot, Incubation, IF-P, Staining, Immunohistochemistry, Wilms Tumor Assay

Journal: Journal of Translational Medicine

Article Title: Mesothelin-based CAR-T cells exhibit potent antitumor activity against ovarian cancer

doi: 10.1186/s12967-024-05174-y

Figure Lengend Snippet: MSLN-CAR T cells can efficiently lyse MUC16-positive human ovarian cancer cells. A Schematic illustration of a CD19-CAR and a MSLN-CAR T vector, respectively. B , C Quantitative analysis of MUC16 protein expression levels in ovarian cells (SKOV3, OVCAR3) by Western blotting ( B ) and flow cytometry ( C ). Tubulin was used as a loading control. Lane 1 = molecular marker; Lanes 2 = SKOV3 cell lysate; Lanes 3 = OVCAR3 cell lysate; Lanes 4 = MUC16 overexpression in 293T cell lysate. B and C consist of three technical replicates. The pictures represent one example of three technical replicates. D The cytotoxicity of MSLN-CAR T cells on ovarian cancer cell lines was quantified using a luciferase assay. Primary human T cells transduced with the indicated lentiviruses were co-incubated with target cells expressing luciferase at an effector to target (E:T) ratio of 2:1 for 20 h. Three independent experiments were performed. E Quantification of IFN-γ ( left ) and TNF-α ( right ) release in response to coculture with CD19-CAR or MSLN-CAR T cells at an E:T ratio of 2:1, as measured by ELISA. Data are presented as the mean ± SD , n = 3. F CAR T cells were co-incubated with target cells expressing luciferase at varying effector to target (E:T) ratios for 20 h. D – F consist of three biologic replicates and three technical replicates. Statistics: two-tailed one-way ANOVA. The results are presented as the mean volume ± SD; *P < 0.05, **P < 0.01, ***P < 0.001 vs CD19-CAR or NTD

Article Snippet: For CSCs, prior to IF staining experiments, such cells were adhered to plates overnight using laminin (Gibco, USA), following which cells were subsequently incubated with anti-MUC16 antibody (sc-365002, Santa Cruz Biotechnology, USA) for one hour followed by washes and fixation with 4% PFA for 5 min.

Techniques: Plasmid Preparation, Expressing, Western Blot, Flow Cytometry, Control, Marker, Over Expression, Luciferase, Transduction, Incubation, Enzyme-linked Immunosorbent Assay, Two Tailed Test

Journal: Journal of Translational Medicine

Article Title: Mesothelin-based CAR-T cells exhibit potent antitumor activity against ovarian cancer

doi: 10.1186/s12967-024-05174-y

Figure Lengend Snippet: MSLN-CAR T cells target MUC16-overexpressing cells for cytotoxic killing. A Structure of the pLGNe-MUC16 expression plasmid used for overexpression assays. B The level of MUC16 protein overexpression in SKOV3 cell line ( left ) was evaluated by Western blot ( light ), with Tubulin was used as a loading control. Lane 1 = molecular marker; Lanes 2 = SKOV3 cell lysate; Lanes 3 = MUC16 overexpression in SKOV3 cell lysate. B and C consist of three technical replicates. The pictures represent one example of three technical replicates. C Flow cytometry was used to detect MUC16 expression on the surface of viable, SKOV3 cells. D Cytotoxicity of MSLN-CAR T cells following their co-culture with MUC16-overexpressing cells as tested with a luciferase-based assay. CAR T cells were incubated with luciferase-expressing target cells for 20 h at an effector to target (E:T) ratio of 2:1. E , F ELISA-based quantification of IFN-γ ( E ) and TNF-α ( F ) released in response to coculture with Mock or MSLN-CAR T cells at an E:T ratio of 2:1. G , H CAR T cells were co-incubated with target cells expressing luciferase at varying effector to target (E:T) ratios for 20 h. D – H consist of three biologic replicates and three technical replicates. Statistics: two-tailed one-way ANOVA. The results are presented as the mean volume ± SD; ns = not significant; *P < 0.05, **P < 0.01 vs CD19-CAR or NTD

Article Snippet: For CSCs, prior to IF staining experiments, such cells were adhered to plates overnight using laminin (Gibco, USA), following which cells were subsequently incubated with anti-MUC16 antibody (sc-365002, Santa Cruz Biotechnology, USA) for one hour followed by washes and fixation with 4% PFA for 5 min.

Techniques: Expressing, Plasmid Preparation, Over Expression, Western Blot, Control, Marker, Flow Cytometry, Co-Culture Assay, Luciferase, Incubation, Enzyme-linked Immunosorbent Assay, Two Tailed Test