Dulbecco s modified eagle s medium ham s f 12 medium

SHSY-5Y cells and HEK293 cells were purchased from Riken Cell Bank (Tsukuba, Japan). Human umbilical vein endothelial cells (HUVECs) were obtained from American Type Culture Collection (ATCC, Manassas, Virginia). SHSY-5Y cells were maintained in Dulbecco’s modified Eagle’s medium/Ham’s F-12 medium (FUJIFILM Wako, Osaka, Japan). HEK293 cells were cultured in Dulbecco’s modified Eagle’s medium (FUJIFILM Wako, Osaka, Japan). The medium for SHSY-5Y and HEK293 cells were supplemented with 10% fetal bovine serum, penicillin (100 units/mL), and streptomycin (100 μg/mL). HUVECs were cultured as previously described [8 (link)]. Cell cultures were maintained in a humidified atmosphere containing 5% CO₂ at 37 °C in the dark. Cells were seeded in glass-bottomed culture dishes 1–2 days before imaging.

MCF‐7 cells (a human mammary carcinoma cell line) [50, 51] and SW‐13 cells (a human adenocarcinoma cell line derived from the adrenal cortex) [52, 53] were purchased from the Japanese Collection of Research Bioresources Cell Bank (Ibaraki City, Japan). The cells were grown in Dulbecco's modified Eagle’s medium/Ham’s F‐12 medium (Fujifilm Wako Pure Chemical, Osaka, Japan) supplemented with 10% fetal calf serum in a 5% CO₂ atmosphere at 37 °C. SW‐13 cells expressing only vimentin as an endogenous IF protein were cloned [53], confirmed by SDS/PAGE and western blotting, and used for subsequent experiments. Transfection of MCF‐7 and SW‐13 cells with plasmids was performed by lipofection [59] using ScreenFect A Plus (Fujifilm Wako Pure Chemical), according to the manufacturer's protocol.

SH-SY5Y cells were purchased from the Riken Cell Bank (Tsukuba, Japan) and were maintained in Dulbecco’s modified Eagle’s medium/Ham’s F-12 medium (FUJIFILM Wako, Osaka, Japan) containing 10% fetal bovine serum, penicillin (100 units/ml), and streptomycin (100 μg/ml). To obtain fluorescence images, cultured cells were seeded onto glass-bottom culture dishes (MatTek Corporation, Ashland, OR, USA).