Journal: ACS Omega

Article Title: Impact of Environmental Pollutant Cadmium on the Establishment of a Cancer Stem Cell Population in Breast and Hepatic Cancer

doi: 10.1021/acsomega.6b00181

Figure Lengend Snippet: Schematic illustration of the monitoring of cadmium-induced CSCs based on the concurrent identification of multiple CSC markers using QD and AOTF-based cellular imaging. MCF-7 and HepG2 cells were treated with cadmium for 72 h, followed by the separation of CSCs using a magnetic sorter. Then, hepatic CSCs were identified based on the expression of QD-conjugated CD133, CD44, and ALDH1, and breast CSCs were identified based on the expression of CD44, CD24, and ALDH1. The AOTF scans the cellular images at a single wavelength emitted from the QD-based nanoprobes without spectral overlap.

Article Snippet: Antibodies specific for CSC markers CD133 (ab-19898; Abcam, USA), CD44 (sc-65265), CD24 (sc-19585), and ALDH1 (sc-374149) (Santa Cruz biotechnology, Santa Cruz, USA) were conjugated to QD705, QD525, QD565, and QD625, respectively, according to the manufacturer’s instructions (Quantum Dot Conjugation Kit; Invitrogen, Carlsbad, CA, USA).

Techniques: Imaging, Expressing

Journal: ACS Omega

Article Title: Impact of Environmental Pollutant Cadmium on the Establishment of a Cancer Stem Cell Population in Breast and Hepatic Cancer

doi: 10.1021/acsomega.6b00181

Figure Lengend Snippet: (a) Breast CSCs obtained from the concurrent monitoring of CD44, CD24, and ALDH1 in the cadmium-treated MCF-7 cell culture. The emission wavelength of QD-conjugated CD44, CD24, and ALDH1 antibodies was measured at 525, 565, and 625 nm, respectively. (b) Quantitative determination of breast CSCs in the MCF-7 cell culture treated at the functional concentration of cadmium. (c) Semiquantitative RT-PCR analysis of CD44, CD24, and ALDH1 expression in MCF-7 cells treated with and without cadmium (1 μM) over the period of 72 h. The values are means ± SD from three independent experiments.

Article Snippet: Antibodies specific for CSC markers CD133 (ab-19898; Abcam, USA), CD44 (sc-65265), CD24 (sc-19585), and ALDH1 (sc-374149) (Santa Cruz biotechnology, Santa Cruz, USA) were conjugated to QD705, QD525, QD565, and QD625, respectively, according to the manufacturer’s instructions (Quantum Dot Conjugation Kit; Invitrogen, Carlsbad, CA, USA).

Techniques: Cell Culture, Functional Assay, Concentration Assay, Reverse Transcription Polymerase Chain Reaction, Expressing

Journal: ACS Omega

Article Title: Impact of Environmental Pollutant Cadmium on the Establishment of a Cancer Stem Cell Population in Breast and Hepatic Cancer

doi: 10.1021/acsomega.6b00181

Figure Lengend Snippet: (a–e) FACS analysis for the breast CSC population induced by cadmium in the MCF-7 cell culture. The x -axis represents CD44 emission, and the y -axis represents ALDH1 emission. For the x -axis, the right side indicates CD44 positive; for the y -axis, the upper side indicates ALDH1 positive. (f) Quantitative determination of breast CSCs obtained using flow cytometry.

Article Snippet: Antibodies specific for CSC markers CD133 (ab-19898; Abcam, USA), CD44 (sc-65265), CD24 (sc-19585), and ALDH1 (sc-374149) (Santa Cruz biotechnology, Santa Cruz, USA) were conjugated to QD705, QD525, QD565, and QD625, respectively, according to the manufacturer’s instructions (Quantum Dot Conjugation Kit; Invitrogen, Carlsbad, CA, USA).

Techniques: Cell Culture, Flow Cytometry

Journal: ACS Omega

Article Title: Impact of Environmental Pollutant Cadmium on the Establishment of a Cancer Stem Cell Population in Breast and Hepatic Cancer

doi: 10.1021/acsomega.6b00181

Figure Lengend Snippet: Gene expression of CSC markers in HepG2 cells and MCF-7 cells treated with 25 μM Ras inhibitor and 1 μM cadmium over the period of 72 h. The mRNA expression was analyzed by RT-PCR. (a) mRNA expression of CD44 and ALDH1 in MCF-7 cells. (b) mRNA expression of CD44, CD133, and ALDH1 in HepG2 cells. Values are means ± SD from three independent experiments.

Article Snippet: Antibodies specific for CSC markers CD133 (ab-19898; Abcam, USA), CD44 (sc-65265), CD24 (sc-19585), and ALDH1 (sc-374149) (Santa Cruz biotechnology, Santa Cruz, USA) were conjugated to QD705, QD525, QD565, and QD625, respectively, according to the manufacturer’s instructions (Quantum Dot Conjugation Kit; Invitrogen, Carlsbad, CA, USA).

Techniques: Gene Expression, Expressing, Reverse Transcription Polymerase Chain Reaction

Journal: Frontiers in Immunology

Article Title: Phenotypic and Functional Analyses Guiding Combination Immune Checkpoint Immunotherapeutic Strategies in HTLV-1 Infection

doi: 10.3389/fimmu.2021.608890

Figure Lengend Snippet: Expression of NCR ligands on dendritic cells in HTLV-1 infection. Cryopreserved PBMCs from SC, AC and HAM/TSP were immunophenotyped for assessment of NCR ligand expression on two dendritic cell (DC) lineages, myeloid DC (mDCs) (A, C, E) and plasmacytoid DC (pDCs) (B, D, F) . mDCs were defined as Lin- (CD3-CD14-CD7-CD19-CD20-) CD11c+CD123- cells; pDCs were defined as Lin- CD11c- CD123+ cells. PD-L1 (PD-1 ligand) (A,B) and PVR (TIGIT ligand) (C,D) single expression and co-expression (E,F) are shown. Mann-Whitney tests were used to determine statistical differences between groups. P -values < 0.05 considered to be significant.

Article Snippet: The following mAbs were used in various panels: from BD Biosciences (San Jose, California) Brilliant Violet 510-conjugated anti-CD4 (OKT4), Flourescein isothiocyanate (FITC)-conjugated anti-CD8 (HIT8a), Phycoerythrin (PE)-conjugated anti-CD151 (14A2.H1), PE-Cy7-conjugated anti-CD19 (SJ25C1), PE-Cy7-conjugated anti-CD20 (2H7), Qdot 605-conjugated anti-CD8, APC-conjugated anti-CD57 (HCD57), V450-conjugated anti-CD45RA (HI100), PerCP-Cy5.5-conjugated anti-CD3 (SK7), PE-conjugated anti-PVR (SKII.4), PE-Cy7-conjugated anti-CD7 (6B7), APC-conjugated anti-HLA-DR (G46-6), FITC-conjugated anti-Ki67 (35/Ki67); from BioLegend (San Diego, CA), Brilliant Violet 711-conjugate anti-CD3 (OKT3), Brilliant Violet 605-conjugated anti-CD14 (M5E2), PerCP-eFluor 710-conjugated anti-TIGIT (MBSA43), APC-Cy7-conjugated anti-PD-1, Alexa Fluor 700-conjugated anti-CD4 (RPA-T4), Alexa Fluor 647-conjugated anti-CCR7 (G043H7), Brilliant Violet 421-conjugated anti-PD-L1 (29E.2A3), Brilliant Violet 510-conjugated anti-CD11b (ICRF44), Brilliant Violet 605-conjugated anti-CD14 (M5E2), Brilliant Violet 711-conjugated anti-CD16 (3G8), FITC-conjugated anti-CD123 (7G3); from Invitrogen/eBioscience (San Diego, CA), Super Bright 645-conjugated anti-LAG-3 (3DS223H), PE-Cy7-conjugated anti-CD28 (CD28.2), FITC-conjugated anti-LAG-3 (3DS223H), Alexa Fluor 700-conjugated CD11c (3.9); from R&D Systems (Minneapolis, MN), PE-conjugated anti-TIM-3 (344823); from Beckman Coulter (Fullerton, CA), ECD-conjugated anti-CD3 (UCHT1).

Techniques: Expressing, Infection, MANN-WHITNEY