Sk br 3 cells

MDA-MB-231, MCF7, and SKBR3 cells (American Type Culture Collection, ATCC), authenticated by isoenzyme and short tandem repeat (STR) analyses), as well as JIMT1 cells (German Collection of Microorganisms and Cell Culture (Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH), DSMZ; authenticated by STR analyses) were maintained in Gibco DMEM (Life Technologies) supplemented with 10% fetal bovine serum (FBS) and confirmed to be mycoplasma negative. SKBR3 cells (ATCC) stably transfected with hrGFP-LC3B⁶⁵ or MDA-MB-231 cells stably transfected with mRFP-eGFP-LC3B were grown in DMEM supplemented with 10% FBS, 20 mM HEPES, and 1X non-essential amino acids (Gibco) with Geneticin (G418). All cells were maintained at 37 °C with 5% CO₂ and 95% humidity.

TMNK-1 cells (an immortalized human liver endothelial cell line, JCRB1564), MEG-01 s cells (a human megakaryoblastic leukemia cell line, IFO50473), and THP-1 cells (a human acute monocytic leukemia cell line, JCRB0112) were obtained from the JCRB cell bank (Osaka, Japan). Jurkat cells (a human T lymphocyte cell line, RCB0806) were obtained from the RIKEN BRC. SK-BR-3 cells (an HER2⁺ human breast cancer cell line, ATCC® HTB-30) was obtained from ATCC. Jurkat cells expressing human FcγRIIa or FcγRIIIa with the Nuclear Factor of Activated T cells (NFAT)-driven luciferase reporter (Jurkat/FcγRIIa/NFAT-Luc, Jurkat/ FcγRIIIa/NFAT-Luc) were established previously (14 (link), 15 (link)). TMNK-1 cells were maintained in Medium 200 (Thermo Fisher Scientific, Waltham, MA) supplemented with Low Serum Growth Supplement (Thermo Fisher Scientific) at 37°C in a humidified atmosphere containing 5% CO₂. Other cell lines were maintained in RPMI1640 medium supplemented with 10% FBS at 37°C in a humidified atmosphere containing 5% CO₂. The expression of the FcγRs in the cell lines was measured by FITC-labelled anti-CD64 antibody, anti-CD32 antibody, and anti-CD16 antibody (BD Biosciences; San Jose, CA) using BD FACSCanto II (BD Biosciences).

SKBR3 cells were obtained from ATCC. Biotin and fluorescein conjugated Anti-HER2 Affibody Molecule (HER2-AFF-B, termed: (ZHER2:477)2) was purchased from Affibody AB, Bromma, Sweden. Dulbecco’s Phosphate Buffered Saline (DPBS), Dulbecco’s Modified Eagle’s Medium GlutaMAX^TM with high glucose and pyruvate (DMEM), Fetal Bovine Serum, certified, heat inactivated (FBS), Minimum Essential Medium Non-Essential Amino Acids (NEAA) 100× solution, Phosphate Buffered Saline (PBS) 10× solution, CellStripper, Normal Goat Serum (GS), and Quantum Dot (QD) Qdot 655 Streptavidin Conjugate (Strep-QD) were from Fisher Scientific GmbH, Schwerte, Germany. D-Saccharose, Glycine, biotin free and molecular biology grade Albumin Fraktion V (BSA), and Sodium Cacodylate Trihydrate were from Carl Roth GmbH + Co. KG, Karlsruhe, Germany. Electron microscopy grade Formaldehyde (FA) 16% solution was from Science Services GmbH, Munich, Germany. HPLC grade deionized Water, 0.01% poly-L-lysine (PLL) solution (mol wt 70,000–150,000), Sodium Tetraborate, Boric Acid, and Fibronectin-like Engineered Protein Polymer-plus (FLEP), were from Sigma-Aldrich Chemie Gmbh, Munich, Germany. 35 mm-cell culture dishes with uncoated glass bottom were from Greiner bio-one, Frickenhausen, Germany (CELLviewTM four compartments), and from ibidi, Munich, Germany (μ-Dish, with imprinted 50 or 500 μm relocation grid).