Direct comparisons between phenotype of THP-1, N₂ and Nef.

<p>Surface expressions of CD11c, CD14, CD40,CD80,CD83,CD86,HLA-DR of stable cell lines–Nef and N₂ (control) within 4 weeks were determined via flow cytometry following establishment. The histograms showed a direct comparison of CD11c,CD14, CD40,CD80,CD83,CD86,HLA-DR levels for Nef <i>vs</i> N₂ and THP-1 Blue line, Nef; green line, N2; red line, THP-1. One representative of three performed experiments was presented.</p

Establishment of stable cell lines Nef and N₂.

<p>The THP-1 cell was transfected with pEGFP-<i>nef</i> and pEGFP-N₂ respectively, giving rise to Nef and N₂ following the limiting dilution in the presence of 550 µg/ml of G418 within 4 weeks. (A): The visualized PCR product on an ethidium bromide stained 1.2% agarose gel, without any template as controls; (B): a, THP-1 in light background; b, THP-1 in black background; c, EGFP expression by N₂ in light background; d, EGFP expression by N₂ in black background; e, Nef-EGFP expression by Nef in light background; f, Nef-EGFP expression by Nef in black background. Microscopy of cells in culture at 400X.magnification.</p

Comparative morphology and phenotype between THP-1, N₂ and Nef upon co-stimulation with GM-CSF,IL-4,TNFα and ionomycin.

<p>(A): morphology of the above cell types was observed in light background (upper panel) and black one (lower panel) at magnification of 400X; (B): The histograms showed a direct comparison of CD11c, CD14, CD40,CD80,CD83,CD86,HLA-DR levels for Nef <i>vs</i> N₂ and THP-1,(7-day culture based on “5+2” protocol ); (C): The histograms showed a direct comparison of above surface markers expression levels for Nef <i>vs</i> N₂ and THP-1 (2-day culture based on “0+2” protocol). One representative of three performed experiments was presented.</p