cd133 cells  (Miltenyi Biotec)

Journal: Genes & Diseases

Article Title: Restriction of YWHAB-mediated YAP cytoplasmic retention is a novel mechanism underlying stemness maintenance and chemoresistance in ovarian cancer peritoneal metastasis

doi: 10.1016/j.gendis.2025.101519

Figure Lengend Snippet: Knockdown of YWHAB promotes stemness and chemoresistance in OCPM. (A) The diameter and the number of the spheroids derived from YWHAB-knockdown and control primary CR OCPM cells were tested (one-way ANOVA; n = 3 biological replicates). (B) The mRNA level of CD133 (left) and the ALDH activity (right) in indicated cells were analyzed by quantitative PCR and ALDH activity assay, respectively (one-way ANOVA; n = 3 biological replicates). (C) The proportions of CD133 + ALDH + cells in indicated cells were analyzed by flow cytometry (one-way ANOVA; n = 3 biological replicates). (D) The frequencies of in vitro sphere-forming cells (left) and in vivo tumor-initiating cells (right) in indicated cells were analyzed by limiting dilution assay (extreme limiting dilution analyses, for in vitro assay, n = 50 biological replicates per group, three groups (1, 5, and 10 cells per well); for in vivo assay, n = 10 biological replicates per group, three groups (8, 40, and 200 cells per mice)). (E) The IC 50 values of cisplatin in indicated primary CR OCPM cells were tested ( n = 3 biological replicates). ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001.

Article Snippet: After the spheroid cells reached 60% confluence, CD133 + spheroid cells were separated by magnetic-assisted cell sorting technology (#130-100-857, Miltenyi Biotech) according to the manual.

Techniques: Knockdown, Derivative Assay, Control, Activity Assay, Real-time Polymerase Chain Reaction, Flow Cytometry, In Vitro, In Vivo, Limiting Dilution Assay

Journal: Genes & Diseases

Article Title: Restriction of YWHAB-mediated YAP cytoplasmic retention is a novel mechanism underlying stemness maintenance and chemoresistance in ovarian cancer peritoneal metastasis

doi: 10.1016/j.gendis.2025.101519

Figure Lengend Snippet: The effects of YWHAB knockdown on the stemness were significantly abolished in YAP5SA-overexpressing cells. (A) The diameter and the number of the spheroids derived from YAP5SA-overexpressing and empty-vector control cells transfected with shRNAs against YWHAB and control shRNAs were measured (one-way ANOVA; n = 3 biological replicates). (B) The mRNA level of CD133 (left) and the ALDH activity (right) in indicated cells were determined by quantitative PCR and ALDH activity assay, respectively (one-way ANOVA; n = 3 biological replicates). (C) The proportion of CD133 + ALDH + cells in indicated cells was analyzed by flow cytometry (one-way ANOVA; n = 3 biological replicates). (D) The frequencies of in vitro sphere-forming cells (left) and in vivo tumor-initiating cells (right) in indicated cells were analyzed by limiting dilution assay (extreme limiting dilution analyses; for in vitro assay, n = 50 biological replicates per group, three groups (1, 5, and 10 cells per well); for in vivo assay, n = 10 biological replicates per group, three groups (8, 40, and 200 cells per mice)). (E) The IC 50 values of cisplatin in indicated cells were tested by cell viability assay ( n = 3 biological replicates). The inhibition rate of 100 uM cisplatin in indicated cells was provided. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001.

Article Snippet: After the spheroid cells reached 60% confluence, CD133 + spheroid cells were separated by magnetic-assisted cell sorting technology (#130-100-857, Miltenyi Biotech) according to the manual.

Techniques: Knockdown, Derivative Assay, Plasmid Preparation, Control, Transfection, Activity Assay, Real-time Polymerase Chain Reaction, Flow Cytometry, In Vitro, In Vivo, Limiting Dilution Assay, Viability Assay, Inhibition