Figure 5.
Figure 5. Effects of dominant-negative mutant p38 MAPK on HOXB4 expression. (A) A cDNA expression vector containing a dominant-negative mutant of p38 MAPK was introduced by lipofection into UT-7/TPO cells and stable cell lines were selected. Overexpression of the cDNA and activity of p38 MAPK were monitored by Western blotting. (B) Parental UT-7/TPO cells and DN-p38 MAPK-expressing cells were treated with 10 ng/mL TPO (□) or 100 ng/mL TPO (▨) for 24 hours and total RNA was prepared. HOXB4 level was determined by real-time RT-PCR. The results represent the average ± SD of 3 independent experiments. ▪ represents the basal HOXB4 level in UT-7/TPO cells cultured with GM-CSF. (C) Parental UT-7/TPO cells and 2 clones of DN-p38 MAPK-expressing UT-7/TPO cells were cultured with 10 ng/mL TPO for the indicated time periods. Each point represents the average ± SD of triplicate samples.

Effects of dominant-negative mutant p38 MAPK onHOXB4expression. (A) A cDNA expression vector containing a dominant-negative mutant of p38 MAPK was introduced by lipofection into UT-7/TPO cells and stable cell lines were selected. Overexpression of the cDNA and activity of p38 MAPK were monitored by Western blotting. (B) Parental UT-7/TPO cells and DN-p38 MAPK-expressing cells were treated with 10 ng/mL TPO (□) or 100 ng/mL TPO (▨) for 24 hours and total RNA was prepared. HOXB4 level was determined by real-time RT-PCR. The results represent the average ± SD of 3 independent experiments. ▪ represents the basal HOXB4 level in UT-7/TPO cells cultured with GM-CSF. (C) Parental UT-7/TPO cells and 2 clones of DN-p38 MAPK-expressing UT-7/TPO cells were cultured with 10 ng/mL TPO for the indicated time periods. Each point represents the average ± SD of triplicate samples.

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