A factor stimulating RNA polymerase II purified from Ehrlich ascites tumor cells was found to stimulate alpha-amanitin-sensitive RNA synthesis in nuclei isolated from spleen cells of anemic mice, though less than it stimulated purified RNA polymerase II. The fidelity of the resulting RNA synthesis was monitored by measuring the stimulation of globin mRNA synthesis. Globin mRNA was measured quantitatively by DNA-RNA hybridization by using plasmid DNA containing globin DNA sequences. Results showed that the synthesis of globin mRNA was enhanced in isolated nuclei in the presence of this factor coinciding with an increase of overall alpha-amanitin-sensitive RNA synthesis. Thus, it was concluded that an externally added factor did not stimulate random transcription but meaningful RNA synthesis in isolated nuclei.