American Society of Hematology

Figure 6

$Figure 6. Neddylation inhibition and dexamethasone differentially affect NF-κB translocation in BMDC. (A) Protein analysis by western blot (left) of p65 isoform of NF-κB protein using nuclear (N) and cytosolic (C) fractions of BMDC in the presence or absence of 500 nm MLN4924 or 500 nm dexamethasone and stimulated concurrently with LPS (0.5 µg/mL) for 30 minutes. Plot (right) shows densitometric and statistical analysis of p65 NF-κB presence in the nuclear fraction and cytosolic fraction. Lamin A/C and LDH proteins were analyzed to demonstrate the presence of nuclear and cytosolic fractions, respectively. One representative experiment of 3 is shown. (B) Immunocytochemistry analysis of p65 NF-κB (column 3) localization in BMDC cultured in the presence or absence of MLN4924 and stimulated with LPS (0.5 µg/mL) for 1 hour. Cell nuclei were stained using DAPI (column 1). Cytoplasmic actin was stained with phalloidin (column 2). Merged images of staining (column 4). One representative experiment of 3 is shown. **P < .01.$

Neddylation inhibition and dexamethasone differentially affect NF-κB translocation in BMDC. (A) Protein analysis by western blot (left) of p65 isoform of NF-κB protein using nuclear (N) and cytosolic (C) fractions of BMDC in the presence or absence of 500 nm MLN4924 or 500 nm dexamethasone and stimulated concurrently with LPS (0.5 µg/mL) for 30 minutes. Plot (right) shows densitometric and statistical analysis of p65 NF-κB presence in the nuclear fraction and cytosolic fraction. Lamin A/C and LDH proteins were analyzed to demonstrate the presence of nuclear and cytosolic fractions, respectively. One representative experiment of 3 is shown. (B) Immunocytochemistry analysis of p65 NF-κB (column 3) localization in BMDC cultured in the presence or absence of MLN4924 and stimulated with LPS (0.5 µg/mL) for 1 hour. Cell nuclei were stained using DAPI (column 1). Cytoplasmic actin was stained with phalloidin (column 2). Merged images of staining (column 4). One representative experiment of 3 is shown. **P < .01.

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