Fetal bovine serum (fbs)

DAMI cells were grown in Roswell Park Memorial Institute 1640 medium supplemented with 10% fetal bovine serum (FBS, Pan Biotech) and 1% penicillin/streptomycin (Lonza). On day 0, DAMI cells were differentiated with 100 nM phorbol 12-myristate 13-acetate (PMA, P8139-1MG, Sigma Aldrich) and cultivated for 4, 8, 24, and 48 hours in full medium or for 4, 8, and 48 hours in medium lacking 10% FBS. For transfection, DAMI cells were seeded and differentiated with 100 nM PMA and after 24 hours transfected with pEGFP-Sac1, pEGFP-C389S-Sac1 (a kind gift of Dr Gerry Hammond, Department of Cell Biology, University of Pittsburgh School of Medicine), or pmCherry-PI4P-SidM (a kind gift of Dr Antonella De Matteis, Telethon Institute of Genetics and Medicine Pozzuoli; Department of Molecular Medicine and Medical Biotechnology University of Napoli Federico II) with Lipofectamine 3000 transfection reagent (Thermo Fischer). Twenty-four hours after transfection, the cells were fixed and stained for proteins of interest. HEK293T cells were grown in Dulbecco’s modified Eagle’s medium (Pan Biotech) supplemented with 10% FBS (Pan Biotech) and 1% penicillin/streptomycin (Lonza). Cells were seeded in full medium for 24 hours before transfection with the EGFP-Murine Stem Cell Virus (MSCV) constructs using Lipofectamine 3000 transfection reagent or before lysis for western blotting.

The human aortic vascular smooth muscle cell line T/G HA-VSMC (ATCC CEL-1999) or human embryonic kidney 293T cells (ATCC CRL-3216) were maintained and cultured in Roswell Park Memorial Institute (RPMI) 1640 Medium or Dulbecco's Modified Eagle's medium supplemented with 10% FBS (PAN Biotech, Germany) and 1% penicillin/G- streptomycin sulfate in a 5% CO₂ humidified atmosphere with a constant temperature of 37°C.
The T/G HA-VSMCs proliferation model was established by culturing cells in RPMI 1640 Medium containing 20% FBS and 1% penicillin/G- streptomycin sulfate in a 5% CO₂ humidified atmosphere with a constant temperature of 37°C (28 (link)).

Normal human dermal fibroblasts (NHDFs) (three different donors, n = 3) were commercially purchased (PromoCell, Heidelberg, Germany) and cultivated in low-glucose DMEM (Thermo Fischer Scientific, Waltham, MA, USA), supplemented with 10% FBS and 5 µg/mL human insulin (both, PAN Biotech, Aidenbach, Germany), 1 ng/mL FGF (Pepro Tech, Hamburg, Germany) and 1% penicillin/streptomycin (Gibco, Karlsruhe, Germany). As the control, human biceps tendon cells (HBTCs) were isolated according to Kohler et al. [34 (link)] from one donor (n = 1) (under Ethical Grant No. 18-985-101 of the Ethics Committee of the University of Regensburg). HBTCs culture medium was DMEM/F-12 (Gibco), 10% FBS (PAN Biotech), 1 × MEM Amino Acids Solution (Thermo Fischer Scientific), 1% L-ascorbic acid 2-phosphate (stock 0.05M) (Sigma-Aldrich, St. Louis, MO, USA), and 1% penicillin/streptomycin (Gibco). Both cell types were kept in a humidified incubator at 37 °C and 5% CO₂. Medium was changed three times per week, and cells were used for experiments in passages 2–5.

The BV-2 murine microglial cell line and human embryonic kidney (HEK) 293T cell line were cultured in Dulbecco’s modified Eagle’s medium (DMEM, Gibco, Carlsbad, USA) containing 10% fetal bovine serum (FBS, PAN Biotech and Lonsera, Aidenbach, Germany), 100 U/mL penicillin, and 100 μg/mL streptomycin (Gibco). MES23.5, a dopaminergic neuroblastoma cells, was derived from hybridization of rat embryonic mesencephalon cells and the murine neuroblastoma cell line N18TG2 [22 (link)]. The cell line was cultured in Dulbecco’s modified Eagle’s medium/nutrient mixture F-12 (DMEM/F-12, Gibco) with 5% fetal bovine serum (FBS, PAN Biotech, and Lonsera), 100 U/mL penicillin (Gibco), 100 μg/mL streptomycin (Gibco), and 100× insulin-transferrin-selenium (Gibco). All three cell lines were cultured in an incubator at 37°C in a 5% CO₂ atmosphere.

Mouse macrophage cell line, RAW 264.7 (source – ATCC (ATCC® TIB-71™)) was cultured in complete Roswell Park Memorial Institute-1640 medium (RPMI-1640) (PAN Biotech, Aidenbach, Germany) with penicillin (100 U/mL), Streptomycin (0.1 mg/mL), and 2.0 mM L-Glutamine (Himedia Laboratories Pvt. Ltd., Mumbai, MH, India), 10% heat-inactivated fetal bovine serum (FBS) (PAN Biotech, Aidenbach, Germany) at 37ºC in a sterile incubator with 5% CO₂ and appropriate humidity. Enzyme-free cell dissociation reagent (ZymeFree™; Himedia Laboratories Pvt. Ltd, Mumbai, MH, India) was used to maintain the cells [56 ].
Undifferentiated human leukemia monocytic cell line, THP-1 (source – ATCC (ATCC® TIB-202™)) was maintained in complete RPMI-1640 (PAN Biotech, Aidenbach, Germany) supplemented with Penicillin (100 U/mL), Streptomycin (0.1 mg/mL), and 2.0 mM L-Glutamine (Himedia Laboratories Pvt. Ltd., Mumbai, MH, India), 10% heat-inactivated FBS (PAN Biotech, Aidenbach, Germany) at 37ºC in a sterile incubator with 5% CO₂ and appropriate humidity. THP-1 cells were further treated with 100 ng/ml PMA for 24 h to differentiate monocytes into macrophage-like cells [75 ].

The human oral squamous cell line CAL27 cells were cultured in Dulbecco's modified Eagle's medium (Gibco) supplemented with 10% fetal bovine serum (FBS, PAN Biotech). HUVECs were cultured in endothelial cell basal medium (ECM, ScienceCell) supplemented with 10% FBS and 1% endothelial cell growth supplement (ECGS). The human monocyte line THP‐1 cells were passaged every 3 d in Roswell Park Memorial Institute (RPMI) 1640 (Gibco) medium supplemented with 10% FBS (PAN Biotech). For the biotinylation, cells were cultured and passaged in complete growth medium containing DSPE‐PEG‐Biotin (0–30 µg mL⁻¹) at 37 °C with 5% CO₂ in a humidified atmosphere for indicated time.

MCF-7 (provided by American Type Culture Collection, ATCC, Manassas, Virginia, USA), MDA-MB-436 (provided by University Clinic Ulm, Ulm, Germany), MDA-MB-453 (provided by University Clinic Ulm, Ulm, Germany), MDA-MB-468 (provided by University Clinic Ulm, Ulm, Germany), and ZR75-1 (provided by Experimental Pharmacology and Oncology, Berlin-Buch, Berlin, Germany) were cultured in DMEM with L-glutamine (Gibco/ThermoFisher Scientific, Waltham, MA, USA), 10% FBS (Pan Biotech, Aidenbach, Germany), 1.2% L-glutamine (Gibco/ThermoFisher Scientific), 1.0% Penicillin-Streptomycin-Glutamine (100×) (Gibco/ThermoFisher Scientific), 1.0% MEM NEAA (non essential amino acid) (Gibco/ThermoFisher Scientific), 0.1% human recombinant insulin (Gibco/ThermoFisher Scientific), and 0.1% hEGF (Sigma-Aldrich/Merck, St. Louis, MO, USA) (Table 1) [22 (link),23 (link)]. HCC-1937 cells (provided by ATCC) were cultured in RPMI 1640 medium with 15% FBS (Pan Biotech, Aidenbach, Germany) and 1% of Penicillin-Streptomycin-Glutamine (100X) (Gibco/ThermoFisher Scientific). Cells were cultured in a humid 5% CO₂ incubator at 37 °C and all cell lines were negative for mycoplasma, verified by PCR.