Expression of caspase mRNA and proteins in untreated and CD437-treated NB4 cells. NB4 cells (300,000/mL) were grown in basal conditions (A) and treated for the indicated amount of time (B) with vehicle (DMSO, Control) or with 10−6 mol/L CD437. (A) One microgram of total RNA was reverse transcribed and PCR amplified with couple of amplimers specific for the indicated caspase cDNAs or for actin cDNA. One tenth of the PCR reaction mixture was run on a 1.2% agarose gel stained with ethidium bromide. Appropriate blanks reverse transcribed in the absence of RNA (no RNA) were set up for each RT-PCR reaction; only the blank corresponding to actin is shown. The molecular weight of DNA standards is indicated on the left. (B) Aliquots of the NB4 homogenates containing an equivalent amount of protein were subjected to Western blot analysis with antibodies specific for the indicated caspases or for actin. The molecular weight of protein standards is indicated on the left. The data are representative of those obtained in three separate experiments.
