Cell Culture and Transfection Protocols

293T human embryonic kidney cells, human cancer cell lines (U-87, MCF-7, HeLa, 293T, AN3CA, Mia Paca-2, PANC-1), and early-passage (p3) mouse embryonic fibroblasts (MEFs) were cultured in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with fetal bovine serum (FBS) and antibiotics as described^{10 (link)}. Transfection was carried out by the calcium phosphate-DNA precipitation method^{37 (link)}. CML-derived K562 cells were maintained and the growth curve analysis was performed as described^{10 (link)}. Where indicated, cells were washed in PBS, and re-plated in fresh medium without curcumin and PGV-1. Viable and dead cells were counted using the trypan blue exclusion method. In some cases, cells were fixed on slide glass by cytospin and inspected after staining with May-Grünwald Giemsa solution (Merck).