1. Regulation of serum-induced lipid accumulation in human monocyte-derived macrophages by interferon-γ. Correlations with apolipoprotein E production, lipoprotein lipase activity and LDL receptor-related protein expression 
Brett Garner, Anna Baoutina, Roger T Dean, Wendy Jessup 
Atherosclerosis 
Volume 128, Issue 1, Pages (January 1997)
DOI: /S (96)

2. Fig. 1
Human serum induces MDM lipid accumulation. Purified human monocytes (1×106) were cultured for 7 days in 10% FCS (A) or 10% HS (B). After rinsing with PBS, cellular lipids were dissolved in 0.2 M NaOH and then extracted into hexane and analysed by HPLC with UV210 detection as described in Section 2. The lipid composition of the d<1.006 (TGRL) fraction of pooled non-fasted HS is shown (C) to demonstrate the retention times of the lipid components marked. TG, triglyceride; CE20:4, cholesteryl arachidonate; CE18:2, cholesteryl linoleate; CE18:1, cholesteryl oleate.
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3. Fig. 2
Effect of HS and FCS on MDM apo E production and secreted LPL activity. Purified human monocytes (1×106) were cultured for 7 days in 10% HS (solid bars) or 10% FCS (hatched bars). After a further 18 h in serum free medium, secreted and cell-associated apo E was measured by Western blot techniques and quantitated using a standard of serially diluted human plasma of known apo E concentration and lipase activity was determined as described in Section 2. Data are mean values and S.E.M. (or range where appropriate) from at least 2 independent experiments.
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4. Fig. 3
Inhibition of MDM apo E production, secreted LPL activity and LRP expression by IFNγ. Purified human monocytes (1×106) were cultured for 14 days in 10% HS (solid bars). Some of these cells also had IFNγ present from day 7 in culture (hatched bars) or from day 0 (open bars). After a further 18 h in serum free medium, secreted and cell-associated apo E and LRP expression was measured by Western blot. Lipase activity was determined as described in Section 2. IFNγ was used at a concentration of 500 U/ml (apo E and LPL) or 1000 U/ml (LRP). Values are expressed as a percentage of the control cell (not exposed to IFNγ at any time) values. Data are representative of at least 2 independent experiments.
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5. Fig. 4
Dose dependence of IFN-γ mediated inhibition of MDM apo E production and LRP expression. Purified human monocytes (1×106) were cultured for 14 days in 10% HS. Some of these cells also had IFNγ present at concentrations of 100, 500 and 1000 U/ml between days 7–14. At day 14, cell-associated (A) and secreted apo E (B) and LRP expression (C) was measured by Western blot. The alkaline phosphatase detection system was used in (A) whereas the HRP-ECL detection system was used in (B) and (C) as described in Section 2. The positions of molecular weight markers are shown by arrows (units are kDa). Dose response curves for the inhibitory effects of IFNγ are also shown (D). Cell-associated apo E (squares); secreted apo E (circles); LRP (triangles). The units for the dose response curves are relative integrated optical densities of the bands shown in A–C, where the culture conditions in the absence of IFNγ are defined as 1.
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6. Fig. 5
Treatment of lipid loaded MDM with IFNγ (500 U/ml) induces morphological alterations. Purified human monocytes were cultured for 14 days in 10% HS alone (A) or for 7 days in 10% HS plus a further 7 days in 10% HS in the presence of IFNγ (500 U/ml) (B). Cells were viewed in situ in plastic tissue culture wells using an inverted microscope. Arrows indicate elongated cellular processes. Original magnification×200.
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7. Fig. 6
IFNγ inhibits MDM lipid accumulation induced by HS. Purified human monocytes (1×106/well) were cultured for 7 days in 10% HS (A) or in 10% HS plus IFNγ (500 U/ml) (B). Lipids were analysed by HPLC as described in the legend to Fig. 1.
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8. Fig. 7
Culture of MDM in 10% HS produces oil red-O positive, lipid loaded cells and is inhibited by IFNγ. Purified human monocytes (1×106) were cultured for 3 (A) or 7 (B) days in 10% HS alone, or with 10% HS plus IFNγ (500 U/ml) for 3 (C) or 7 days (D). After washing with PBS, cells were stained with oil red-O in situ in tissue culture plates and viewed using oil immersion microscopy. Original magnification×1250.
Atherosclerosis  , 47-58DOI: ( /S (96) )