1. Volume 25, Issue 1, Pages 197-207 (January 2017)
High-Density Lipoproteins Exert Pro-inflammatory Effects on Macrophages via Passive Cholesterol Depletion and PKC-NF-κB/STAT1-IRF1 Signaling 
Emiel P.C. van der Vorst, Kosta Theodorou, Yongzheng Wu, Marten A. Hoeksema, Pieter Goossens, Christina A. Bursill, Taghi Aliyev, Leonie F.A. Huitema, Sander W. Tas, Ine M.J. Wolfs, Marijke J.E. Kuijpers, Marion J. Gijbels, Casper G. Schalkwijk, Debby P.Y. Koonen, Shahla Abdollahi-Roodsaz, Kimberly McDaniels, Chih-Chieh Wang, Michael Leitges, Toby Lawrence, Jogchum Plat, Miranda Van Eck, Kerry-Anne Rye, Lhousseine Touqui, Menno P.J. de Winther, Erik A.L. Biessen, Marjo M.P.C. Donners 
Cell Metabolism 
Volume 25, Issue 1, Pages (January 2017)
DOI: /j.cmet
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2. Cell Metabolism 2017 25, 197-207DOI: (10.1016/j.cmet.2016.10.013)
Copyright © 2017 Elsevier Inc. Terms and Conditions

3. Figure 1
Pro-inflammatory Functions of HDL on Murine and Human Macrophages In Vitro and In Vivo
(A) Cytokine secretion by native HDL-treated and LPS-stimulated BMDMs is shown.
(B) Cytokine secretion by 300 μg/ml native HDL-treated BMDMs using different LPS incubation times is shown.
(C) Cytokine secretion by 300 μg/ml native HDL-treated and LPS-stimulated BMDMs using different HDL incubation times is shown.
(D) In vivo HDL priming results in more inflammatory, LPS-triggered PMs re-stimulated in vitro with LPS (n = 6).
(E) TNF-α cytokine secretion by HDL-treated WT BMDMs, stimulated with various TLR ligands, is shown.
(F) In vivo HDL priming results in more inflammatory, R848-triggered PMs re-stimulated in vitro with R848 (n = 6).
(G) In vitro P. aeruginosa phagocytosis by alveolar macrophages (MHS cell line) is shown. Representative image is of n = 2.
(H) Bacterial titers in BALs are shown after P. aeruginosa infection over time.
(I) HE-stained lung cross-sections after infection are shown.
(J) BAL cell contents after P. aeruginosa infection are expressed as a ratio. Note the difference in the y axis, reflecting neutrophil influx over time. Statistical analysis uses Kruskal-Wallis with Dunn’s post-test.
(K) BAL fluid cytokine profile is shown after P. aeruginosa infection. Note the difference in the y axis.
All results are expressed as mean ± SEM; n.d., not detectable. See also Figures S1 and S2.
Cell Metabolism  , DOI: ( /j.cmet )
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4. Figure 2
Lipid Raft Disruption by Cholesterol Depletion Mediates Pro-inflammatory HDL Effects
(A) Cytokine secretion by HDL-treated and LPS-stimulated BMDMs is shown. The dotted line represents HDL’s effect in WT BMDMs.
(B) Lipid raft staining of WT BMDMs or WT and Tg PMs and representative pictures of BMDMs are shown.
(C) Cytokine secretion by cholesterol depleted and LPS-stimulated BMDMs is shown. Additionally, BMDMs were pre-treated with MβCD or a MβCD and cholesterol mixture and washed before HDL incubation.
(D and E) Cytokine secretion by HDL-treated and LPS-stimulated BMDMs, with or without ADAM inhibition (D) or soluble TNFR Enbrel (E), is shown.
All results are expressed as mean ± SEM.
Cell Metabolism  , DOI: ( /j.cmet )
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5. Figure 3
Macrophage Pro-inflammatory HDL Effects Are Mediated by NF-κB Activation and STAT1/IRF1 Signaling
(A) De novo motif analysis of LPS-induced genes further upregulated by HDL is shown.
(B and C) Microarray expression levels of NF-κB-p65 (B) and IRF1-responsive genes (C) are represented as heatmaps. Red and green indicate increased and decreased expression, respectively, compared to average LPS conditions.
(D) Cytokine secretion by HDL-treated and LPS-stimulated BMDMs is shown.
(E) NF-κB luciferase levels in HDL-treated BMDMs after 3 hr LPS stimulation are shown.
(F) p65-ChIP analysis of treated BMDMs after 3 hr LPS stimulation is shown.
(G) Cytokine secretion by HDL-treated and LPS-stimulated and IKKα- and IKKβ-inhibited BMDMs is shown.
All results are expressed as mean ± SEM. See also Figure S3.
Cell Metabolism  , DOI: ( /j.cmet )
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6. Figure 4 PKC Mediates HDL’s Pro-inflammatory Effects in Macrophages
(A) PKC activity in BMDMs incubated with nHDL or Ro is shown.
(B–F) Cytokine secretion by HDL-treated and LPS-stimulated BMDMs is shown with or without Ro (B), UCN-01 (C), Gö6976 (D), PKCδ, PKCε, and PKCθ inhibitor (E), or both Gö6976 and PKCδ, PKCε, and PKCθ inhibitor (F).
(G) Proposed (simplified) mechanism of pro-inflammatory action of HDL in macrophages is shown.
All results are expressed as mean ± SEM. See also Figure S4 and Table S1.
Cell Metabolism  , DOI: ( /j.cmet )
Copyright © 2017 Elsevier Inc. Terms and Conditions