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: iScience

Article Title: Preoperative immune checkpoint inhibition and cryoablation in early-stage breast cancer

doi: 10.1016/j.isci.2024.108880

Figure Lengend Snippet:

Article Snippet: Briefly, one million PBMCs and TILs were washed with 2 mL FACS buffer (PBS [phosphate-buffered saline] containing bovine 1% serum albumin and 0.05 mM EDTA), resuspended in 50 μL FACS buffer and stained with a fixable Aqua viability dye (Invitrogen) and a cocktail of antibodies to the following surface markers: CD8-Qdot 605 (Invitrogen, 3B5), CD4-Qdot 655 (Invitrogen, S3.5), PD-1-PE (BD, MIH4), LAG-3-FITC (Enzo Life Sciences, 17B4), ICOS-PE-Cy7 (eBioscience, ISA-3), TIM-3-APC (R&D Systems, 344823).

Techniques: Recombinant, Saline, Staining, Extraction, Software, Flow Cytometry

Journal: Scientific Reports

Article Title: IL-6 ameliorates acute lung injury in influenza virus infection

doi: 10.1038/srep43829

Figure Lengend Snippet: ( a ) Immunohistochemical examination of NP-positive cells in thioglycollate-elicited macrophages infected with IAV at an MOI of 1 for 24 h. Representative images are shown (original magnification ×100, scale bar = 200 μm). ( b ) NP-positive macrophages were quantified by averaging the number of positively stained cells in four randomly selected fields ( n = 3). ( c ) Titers of IAV produced from the infected macrophages were quantified by the plaque assay ( n = 3). ( d ) Immunohistochemical detection of macrophages in the lung with anti-Mac-3 antibody at day 7 p.i. in WT and IL-6 −/− mice infected with 10 5 PFU of IAV. Macrophages in the lung were quantified by averaging the number of Mac-3-positive cells in three randomly selected fields in each section ( n = 4). ( e ) Immunohistochemical detection of macrophages in the BAL fluid with anti-F4/80 antibody at day 7 p.i. in WT and IL-6 −/− mice infected with 10 5 PFU of IAV. Total numbers of macrophages in the BAL fluid were quantified by counting F4/80-positive cells ( n = 4). ( f ). Total numbers of thioglycollate-elicited peritoneal macrophages collected from WT and IL-6 −/− mice were counted using a hemocytometer ( n = 3 for WT, n = 5 for IL-6 −/− ). ( g ) Migratory capability of thioglycollate-elicited peritoneal macrophages from WT and IL-6 −/− mice determined by the Boyden chamber assay. The number of migrating cells was the average of the cells counted from four randomly selected fields in each section ( n = 6). ( h ) Cytotoxicity of thioglycollate-elicited peritoneal macrophages from WT and IL-6 −/− mice infected with IAV at an MOI of 1 for 24 h, as assessed by the LDH release assay ( n = 4).

Article Snippet: To assess the phagocytosis of viruses or nanoparticles by macrophages, peritoneal macrophages (10 6 cells) were incubated with IAV that had been labeled with FITC (NHS-Fluorescein; Pierce, Rockford, Ill) at an MOI of 1 as described previously or with 5 × 10 10 QD649 quantum dot particles for 30 min at 37 °C and then treated with 40 μl of 0.1% trypan blue to quench extracellular florescence.

Techniques: Immunohistochemical staining, Infection, Staining, Produced, Plaque Assay, Boyden Chamber Assay, Lactate Dehydrogenase Assay