Fig. 6

Mephedrone activates NF-κB signaling pathway. a TransAM Transcription Factor Activation Assay was used to measure concentrations of activated NF-κB subunits in the nuclear fractions of endothelial cells treated with 10 μM mephedrone for 24 h. Mephedrone significantly upregulates the level of nuclear p65 subunit. Data shown as mean ± SEM. Experiments were independently performed three times. Within each individual experimental set, primary cells from three different donors were used (n = 9). b Representative image of p65 nuclear translocation in endothelial cells. Scale bar is 10 μm. Representative immunoblot (c) and densitometry analysis (d) of TLR4 expression in cytosolic and membrane fractions of hBMVEC. Data shown as mean ± SEM. Experiments were independently performed three times. Within each individual experimental set, primary cells from three different donors were used (n = 9). e TransAM NF-κB p65 Activation Assay was used to measure concentrations of activated p65 in the nuclear fraction of endothelial cells treated with 10 μM mephedrone alone or pre-treated with 100 nM TAK242 for 3 h. TAK242 significantly downregulates the amount of mephedrone-activated p65 subunit. Data shown as mean ± SEM. Experiments were independently performed three times. Within each individual experimental set, primary cells from three different donors were used (n = 9)