Dulbecco s phosphate buffered saline dpbs 1x

Tumor cells were grown to 70% confluence, washed with Dulbecco’s phosphate-buffered saline (DPBS) 1X (Biowest) and stained in a flask with lipophilic dyes—Vybrant CM-DiI (4 μl/ml in DPBS 1X), green CMFDA (1 μl/ml in DPBS 1X, 1 mM stock), or Deep Red Cell Tracker (1 μl/ml in DPBS 1X, 10 mM stock) (Life Technologies), for 10 min at 37 °C, in darkness. Cells were washed with DPBS and detached with 2 mM EDTA by scrapping. Cell suspension was collected to 1.5 ml eppendorfs, centrifuged at 250 × g, for 4 min at 4 °C, and resuspended in DMEM. Cell viability was assessed by trypan blue exclusion method, and cell number was determined by hemocytometer counting. Cells were resuspended in DPBS 1X to a final concentration of 0.25 × 10⁶ cells/μl.

The 293T cells were washed with Dulbecco’s phosphate buffered saline (DPBS, 1X; Biowest), counted and incubated overnight with different IDLVs. The media were washed and the cells were exposed to 1, 4, and 8 mM of etoposide for 2 or 8 h. The cells were then washed and eGFP expression levels were studied by flow cytometry over time.