fig4

Figure 4. Nef increases intra-cellular and intra-nuclear Dox by LRP downregulation. A549 or A549/Dox cells were treated with 10 μM Nef and/or 3 μM Dox and (A) and (B). Analysed for LRP expression, representative western blot image of A549 and A549/Dox cells treated with Dox alone, Nef alone and Nef and Dox combination and densitometric analysis are shown. A549 or A549/Dox cells were knocked out for LRP using siLRP and treated with Dox alone or in combination with Nef; C: the intracellular Dox was assessed and the results were expressed as nmoles of Dox; D: cell vaibility was assessed; and E: intranuclear Dox accumulation was assessed. The results shown are mean ± SEM and *** represents P < 0.01 or ** represents P < 0.05, which are three separate experiments performed in triplicate. Nef: Neferine; Dox: Doxorubicin; LRP: lung resistance protein; siLRP: small interfering LRP; GAPDH: Glyceraldehyde 3-phosphate dehydrogenase