The β-chain, GATA-1, and FOG-1 mRNA levels in Ba/F3 and PT18. (A) RT-PCR was performed to measure FcϵRI β-chain, FOG-1, GATA-1, and GAPDH mRNA levels, as visualized by ethidium bromide staining of agarose gels. (B) Western blotting analyses of FOG-1 and GATA-1. Whole-cell lysates (1 × 106 cells per lane) were applied into each well and were analyzed with anti-FOG-1 or anti-GATA-1 Ab as the primary Abs followed by peroxidase-conjugated anti-goat IgG donkey Ab or peroxidase-conjugated anti-rat IgG rabbit Ab as the secondary Abs, respectively. (C) Transcription activities of the β-chain and αIIb promoters. Five micrograms of each reporter plasmid was introduced into Ba/F3 or PT18. The relative luciferase activity driven by β-69/pGL3-Basic (β-69; ▪) or αIIb/pGL3-Basic (αIIb; ▦)is represented as the ratio to the activity driven by pGL3-Basic (Basic; □). Each experiment was conducted in duplicate for each sample, and the results are expressed as mean ± SD for more than 3 independent experiments in Figures 1, 2, 3, and 5.
