Fig. 5.
Fig. 5. Overexpression of p27KIP1 induces apoptosis in DCs. / (A) Evaluation transduction efficiency of rAd-LacZ or rAd-p27–infected immature DCs using X-gal. (B) DCs were infected with rAd-LacZ or rAd-p27. After 48 hours of culture in the presence of IL-4 and GM-CSF, infected DCs and control DCs were harvested for the preparation of whole cell lysates. Then equal amounts of protein (20 μg/lane) were loaded, and the levels of p27KIP1 were determined. Data shown are representative for 3 independent experiments with different donors. (C) DCs were infected with rAd-LacZ or rAd-p27 and subsequently cultured in IL-4 and GM-CSF. After 48 hours of culture, annexin V–FITC/PI staining was performed to determine the percentage of apoptotic cells. Data shown are representative for 2 independent experiments with different donors. (D) DCs were left untreated or infected with rAd-LacZ or rAd-p27 and subsequently cultured in IL-4 and GM-CSF. Cells were harvested after 24, 48, and 72 hours of culture. Annexin V–FITC/PI stainings were performed to determine the percentage of annexin V+/PI− (early apoptotic) cells (n = 2).

Overexpression of p27KIP1 induces apoptosis in DCs.

(A) Evaluation transduction efficiency of rAd-LacZ or rAd-p27–infected immature DCs using X-gal. (B) DCs were infected with rAd-LacZ or rAd-p27. After 48 hours of culture in the presence of IL-4 and GM-CSF, infected DCs and control DCs were harvested for the preparation of whole cell lysates. Then equal amounts of protein (20 μg/lane) were loaded, and the levels of p27KIP1 were determined. Data shown are representative for 3 independent experiments with different donors. (C) DCs were infected with rAd-LacZ or rAd-p27 and subsequently cultured in IL-4 and GM-CSF. After 48 hours of culture, annexin V–FITC/PI staining was performed to determine the percentage of apoptotic cells. Data shown are representative for 2 independent experiments with different donors. (D) DCs were left untreated or infected with rAd-LacZ or rAd-p27 and subsequently cultured in IL-4 and GM-CSF. Cells were harvested after 24, 48, and 72 hours of culture. Annexin V–FITC/PI stainings were performed to determine the percentage of annexin V+/PI (early apoptotic) cells (n = 2).

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