Mda 435

DKMG cells were purchased from the DSMZ cell bank (Braunschweig, Germany). Lung cancer (H1299, H1975), myeloid leukemia (K562), breast cancer (MDA-435), normal breast (MCF-10A), and embryonic (HEK-293) cell lines were obtained from the ATCC. All cell lines were cultured in RPMI 1640 or DMEM medium supplemented with 10% fetal bovine serum (FBS), 2 mM L-glutamine, and 1% penicillin/streptomycin (all from Sartorius, Israel). Cells were maintained at 37 °C in humidified 5% CO₂ atmosphere. H1299, H1975, and DKMG cells were characterized by cell surface overexpression of EGFR [32 (link)]. DKMG cells are known to express two forms of EGFR—wild type (WT) and another carrying a mutation in the external region of the receptor (EGFR^vIII) [33 (link)]. H1975 cells carry a mutation in the internal domain of EGFR (L858R/T790M) [34 (link)].

The human breast cancer cell lines BT‐474, MCF‐7, MDA‐231, MDA‐468, the melanoma‐derived cell line, MDA‐435, and the immortalized breast epithelial cell line, MCF‐10A, were purchased from ATCC. MCF‐10A cells were grown in F12/DMEM (50/50) medium, and the rest of the cells were all cultured in RPMI medium. All culture media were supplemented with 10% (v/v) fetal bovine serum (FBS) and 1% (v/v) penicillin/streptomycin antibiotics (ATCC). The human prostate cancer cell line, DU‐145, was purchased from ATCC, and was also maintained in RPMI medium supplemented with 10% FBS and 1% penicillin/streptomycin. All the cells were maintained in a humidified atmosphere at 37°C in incubators with 5% CO₂. The DU‐145 cells were chosen based on our previous studies that showed the cells to be resistant to the activity of both free and encapsulated IPA‐3.18

Human MDA-231, MDA-435, DU145, HT1080, U251, U87F-7, MCF-7 and Hs27 cell lines were from the American Type Culture Collection. MDA-231, MDA-435, and DU145 were maintained in complete RPMI 1640 and HT-1080, U251, U87, MCF-7 and Hs27 cells in DMEM supplemented with 10% fetal bovine serum, 100 IU/mL penicillin, 100 μg/mL streptomycin, 2 mmol/L l-glutamine, and 25 mmol/L HEPES buffer for RPM1 and sodium pyruvate for DMEM, respectively at 37°C in 5% CO₂ incubator. For western blots, cell lysates were prepared, protein concentration was determined using Bradford reagent and equal amounts of cellular protein were loaded. For immunofluorescence, cells grown on glass coverslips were fixed with 3% paraformaldehyde and antibody-labeled as previously described [18 (link)]. Images were collected with ×60 or ×100 planapochromat objectives (numerical aperture, 1.35) of an FV1000 Olympus confocal microscope.