Expression of FL mRNA during aplasia. (A) Semiquantitative RT-PCR analysis in PBMC from patients during chemotherapy treatment. Twenty-five PCR cycles were performed under conditions of the linear accumulation of FL and β-actin PCR products (B). Southern blots were hybridized with 32P-labeled internal probes, and radioactivity in each band was quantified by phosphorimaging analysis. Autoradiography was for 4 hours (FL) and 30 minutes (β-actin). Three analyses were performed for each of 4 analyzed patients; representative results for 2 patients are shown. Days of treatment are indicated above and peripheral leukocyte counts below the autoradiograms. Methods more accurate than RT-PCR could not be applied given the extremely limited availability of patients’ cells at the time of treatment. (C) Semiquantitative RT-PCR analysis in mouse tissues during irradiation-induced aplasia. Mice received 8.0 Gy total body irradiation. RNA was isolated before irradiation (day 0), during aplasia (days 3 and 6), and at the beginning of recovery (day 10), and FL mRNA expression was analyzed as described above. Three analyses were performed for 2 animals; representative results are shown. The BM nucleated cell count per femur plus tibia is indicated below the autoradiogram.
