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Figure 2. Expression of c-fos following Ca2+ entry through CRAC channels. A) RT-polymerase chain reaction (RT-PCR) showing thapsigargin stimulation (4 min) in the presence of external Ca2+ results in c-fos gene expression but not in the absence of external Ca2+ or in the presence of 2-APB. Cells were stimulated with thapsigargin for 4 min and then perfused with Ca2+-free solution for 41 min prior to RNA extraction The upper panel shows control ß-actin loading. B) Western blotting of nuclear membrane shows that Ca2+ entry promotes c-fos protein expression, and this is suppressed by 2-APB (30 µM; 5 min pretreatment). Same stimulation protocol as in (A). C, upper panel) Depicts a typical nuclear membrane blot probing for c-fos expression in nonstimulated cells and after stimulation with thapsigargin in the absence and then presence of U0126. Aggregate data from four gels are summarized in the histogram. D) As in (C) but now in the presence of PD98059.
