Journal: International journal of molecular medicine

Article Title: Autophagy is essential for the endothelial differentiation of breast cancer stem‑like cells.

doi: 10.3892/ijmm.2019.4399

Figure Lengend Snippet: Figure 1. Flow cytometry and microsphere culture of BCSLCs. (A) CD24‑/low cells were isolated from MCF‑7 using immunomagnetic beads, and the expres- sion of CD24 in MCF‑7 and isolated CD24‑/low cells were assessed using flow cytometry. (a) Isotype control for (b); (b) MCF‑7 cells; (c) isotype control for (d); (d) isolated CD24‑/low cells. (B) CD44+ cells were further isolated from CD24‑/low cells and the expression of CD44 was assessed using flow cytometry. (a) Isotype control for (b); (b) CD44+ cells. (C) Expression of CD24 and CD44 in MCF‑7, BCSLCs and BCSLCs after eight passages. (a) isotype control for (b); (b) MCF‑7 cells; (c) CSLCs; (d) isotype control for (e); (e) BCSLCs after eight passages. (D) The isolated BCCSLCs were cultured in microspheres for 0 and 64 h in stem cell culture medium. BCSLC, breast cancer stem‑like cell; PE, phycoerythrin.

Article Snippet: The following primary antibodies were used: Fluorescein isothiocyanate (FITc)-conjugated anti-human cd44 (cat. no. 130-113-903; Miltenyi Biotec GmbH), phycoerythrin (PE)-conjugated anti-human cd24 (cat. no. 130-098-861; Miltenyi Biotec GmbH) PE-conjugated anti-human cd31 (cat. no. 130-110-807; Miltenyi Biotec GmbH) and FITc-conjugated anti-human cd105 (cat. no. 130-098-778; Miltenyi Biotec GmbH).

Techniques: Flow Cytometry, Isolation, Control, Expressing, Cell Culture, Stem Cell Culture

Journal: Genes & Cancer

Article Title: CD133-positive cancer stem cells from Colo205 human colon adenocarcinoma cell line show resistance to chemotherapy and display a specific metabolomic profile

doi:

Figure Lengend Snippet: A-B. Cytometry analyses of Nestin, CK20, CD133, Oct3/4, CD44 and CD44v9 on 10% FBS Colo205 growing cells (control) and serum-free growing cultures (week 1 to week 5).

Article Snippet: The purity of sorted cells was evaluated by flow cytometry using FACSCalibur (BD Biosciences) after labeling with anti-human CD133/2 or CD44 antibody (Miltenyi Biotec).

Techniques: Cytometry, Control

Journal: Genes & Cancer

Article Title: CD133-positive cancer stem cells from Colo205 human colon adenocarcinoma cell line show resistance to chemotherapy and display a specific metabolomic profile

doi:

Figure Lengend Snippet: A. Tumorsphere evaluation of CD133+ and CD133- sorted cells. B. Invasiveness of different cell fractions measured using a collagen-based invasion kit (relative DO measured at 560 nm). C. Survival assay after chemotherapy treatment with cisplatin and 5-FU. D. Metabolite quantification after CE-TOF-MS experiments. Only metabolites of interest for Colo205 cells cultured in 10% FBS (control; green bar), CD133+ sorted cells (red bar) and CD44+ sorted cells (yellow bar) are reported here.

Article Snippet: The purity of sorted cells was evaluated by flow cytometry using FACSCalibur (BD Biosciences) after labeling with anti-human CD133/2 or CD44 antibody (Miltenyi Biotec).

Techniques: Clonogenic Cell Survival Assay, Cell Culture, Control

Journal: Cancer Biology & Therapy

Article Title: Transcription factor 3 promotes migration and invasion potential and maintains cancer stemness by activating ID1 expression in esophageal squamous cell carcinoma

doi: 10.1080/15384047.2023.2246206

Figure Lengend Snippet: TCF3 regulates the expression of cancer stem markers CD44 and CD133 by transcriptionally regulating ID1. a. RNA-seq results show significantly upregulated and downregulated gene signatures. b-c. In KYSE-150 and TE-1, with the knockdown of TCF3 the protein expression level of ID1 is subsequently reduced. d-e. In KYSE-150 and TE-1, with the knockdown of TCF3, the mRNA expression of ID1 is subsequently reduced. f-g. CHIP and Dual luciferase reporter assay suggested TCF3 could transcriptionally regulate ID1.h-i. With knockdown of TCF3 in KYSE-150 and TE-1 CD44, fluorescence intensity decreased. J-K. The protein expression of CD44 and CD133 was reduced with the knockdown of TCF3 in KYSE-150 and TE-1. l -m. With the knockdown of TCF3 in KYSE-150 and TE-1 knockdown increased sensitivity to the chemotherapeutic drug cisplatin. n-o. mRNA expression of ID1 was reduced in KYSE-150 and TE-1 after siRNA knockdown. p-q. The protein expression level of CD44 and CD133 was subsequently reduced in KYSE-150 and TE-1 when ID1 was knocked down. I. KYSE-150 has a significant decrease in the sphere formation efficiency after si-TCF3 was transfected. s. with knockdown of TCF3, CD44+ ESCC cells number were decreased.

Article Snippet: CD44 antibody purchased from R&DSystems (FAB6127G).

Techniques: Expressing, RNA Sequencing Assay, Luciferase, Reporter Assay, Fluorescence, Transfection

Journal: Cancer Biology & Therapy

Article Title: Transcription factor 3 promotes migration and invasion potential and maintains cancer stemness by activating ID1 expression in esophageal squamous cell carcinoma

doi: 10.1080/15384047.2023.2246206

Figure Lengend Snippet: ID1 promotes proliferation, migration, invasion, and drug resistance of ESCC cells. a-b. The fluorescence intensity of ID1 was reduced in KYSE-150 and TE-1 after ID1 was knocked down. c-d. The migration and invasion abilities of KYSE-150 and TE-1 were decreased when ID1 was knocked down. e -f. The wound healing rates of KYSE-150 and TE-1 were decreased when ID1 was knocked down. g-h. Cell proliferation is affected in KYSE-150 and TE-1 after the siRNA knockdown of ID1.I&J. Increased sensitivity to the chemotherapeutic drug cisplatin with knockdown of TCF3 in KYSE-150 and TE-1. k. KYSE150 has a significant decrease in the sphere formation efficiency after si-ID1 was transfected. l. with knockdown of ID1, CD44+ ESCC cells number were decreased.

Article Snippet: CD44 antibody purchased from R&DSystems (FAB6127G).

Techniques: Migration, Fluorescence, Transfection

Journal: Cancer Biology & Therapy

Article Title: Transcription factor 3 promotes migration and invasion potential and maintains cancer stemness by activating ID1 expression in esophageal squamous cell carcinoma

doi: 10.1080/15384047.2023.2246206

Figure Lengend Snippet: The expressions of cancer stem markers CD44 and CD133 were correlated with the progression of ESCC. a. The expression of CD44 is corresponding to the tumor staging of ESCC. b. The expression of CD133 is corresponding to the tumor staging of ESCC. c. The expression of CD44 positively correlates with TCF3 expression.d.CD133 positively correlates with the expression of TCF3. * p < .05, ** p < .01. *** p < .001.

Article Snippet: CD44 antibody purchased from R&DSystems (FAB6127G).

Techniques: Expressing

Journal: International Journal of Molecular Medicine

Article Title: Effects of cisplatin on the proliferation, invasion and apoptosis of breast cancer cells following β-catenin silencing

doi: 10.3892/ijmm.2020.4543

Figure Lengend Snippet: Expression levels of CD44/54 in BC cells are analyzed by flow cytometry. (A) T47D cells treated with cisplatin, siR-β-catenin and the combination of cisplatin and siR-β-catenin were analyzed by FACS and the fluorescence intensities of CD44/CD54 were obtained. (B) Statistical analysis of the expression of CD44/CD54 in T47D cells. (C) The expression of CD44/CD54 in MCF-7 cells treated with cisplatin, siR-β-catenin and the combination of cisplatin and siR-β-catenin were analyzed by FACS. (D) Statistical analysis of the expression of CD44/CD54 in MCF-7 cells. All data are presented as mean ± standard error of the mean. * P<0.05, ** P<0.01 and *** P<0.001 vs. control. BC, breast cancer; siRNA; small interfering RNA; CD44, CD44 antigen; CD54, intercellular adhesion molecule 1.

Article Snippet: The cells were stained with phycoerythrin-labeled mono-clonal CD44 (cat. no. MAB6127; 1:200; R&D Systems, Inc.) or allophycocyanin-labeled CD54 (cat. no. BBA20; 1:300; R&D Systems, Inc.) antibodies or the isotype controls (cat. no. MAB0031; 1:200; R&D Systems, Inc.) at 37°C for 30 min, rinsed twice with PBS and fixed in 10% (v/v) formaldehyde and PBS at 37°C for 25 min. Then, the cells were sorted and observed using a BD FACSCalibur 4-color flow cytometer (BD Biosciences).

Techniques: Expressing, Flow Cytometry, Fluorescence, Small Interfering RNA