Microporator neon transfection system

HeLa cells (obtained from American Type Culture Collection) were maintained in Dulbecco’s Modified Eagle’s Medium (Gibco) supplemented with 10% fetal bovine serum (Invitrogen) at 37 °C in a humidified 10% CO₂ atmosphere. Cells were confirmed to be free from mycoplasma using an e-Myco Mycoplasma PCR detection kit (iNtRON). For live-cell imaging, cells were plated on a 96-well plate (μ-Plate 96 Well ibiTreat; ibidi GmbH) and transfected using either a Microporator (Neon Transfection System; Invitrogen) or Lipofectamine LTX (Invitrogen), according to the manufacturers’ instructions. For HeLa cells, electroporation conditions (two pulses of 980 V for 35 ms) were additionally optimized for higher transfection efficiency.

HeLa cells and HEK293T (ATCC) were maintained in Dulbecco’s Modified Eagle’s Medium (DMEM; PAA Laboratories GmbH) supplemented with 10% fetal bovine serum (Invitrogen) at 37 °C in a humidified 10% CO₂ atmosphere. Cells were transfected using either a Microporator (Neon Transfection System; Invitrogen) or Lipofectamine LTX (Invitrogen) according to the manufacturer’s instructions.