Bcl-2 overexpression fails to protect 32D-Kit cells from SLF plus Ca²⁺ influx blocker. (A) Western blot of cell lysates from 32D-Kit (lane 1) or 32D-Kit-bcl-2 (lane 2) cells. Cell lysates were separated by SDS-PAGE, transferred to nitrocellulose, and probed with anti–bcl-2 antibodies (top panel). The blot was then stripped and reprobed with an anti-actin monoclonal antibody to demonstrate equal protein loading (bottom panel). (B) 32D-Kit (▪) or 32D-Kit-bcl-2 (▨) cells were incubated for 18 hours with SLF or IL-3 plus 2.5, 5, or 10 μmol/L econazole, and the proportion of dead cells was determined by Trypan Blue exclusion. In additional cultures, IL-3 alone or no factor was added and cell viability was similarly determined.