Quantitative RT-PCR and ChIP Assays

All cells and tissues were processed and analyzed as previously described^{5 (link)}. For RT-PCR, total RNA was extracted using a TRIzol extraction method. The sequences of primers for RT-PCR and ChIP assays are listed in Supplementary Tables S2 and S3^{5 (link)}. Flag-tagged MafB was purified with M2 Affinity Gel as previously described^{5 (link)}, and Myc/DDK-tagged LXRα was obtained from OriGene. Immunoblot and ChIP analyses used the following antibodies: anti-β-actin (ab8226; Abcam), anti-GAPDH (sc-25778; Santa Cruz), anti-MafB (MAB3810; R&D Systems), anti-Stat6 (sc-621; Santa Cruz), anti-pStat6 (sc-11762; Santa Cruz), anti-LXRα (ab41902; abcam), anti-Flag (M2) or anti-HA Affinity Gel (Sigma), goat anti-rabbit IgG-HRP (sc-2054; Santa Cruz), IRDye 800CW–conjugated goat anti–mouse and anti–rabbit IgG (926-32210 and 926-32211, respectively; LI-COR), and IRDye 800–conjugated anti-Flag and anti-HA (600-432-383 and 600-432-384, respectively; Rockland). Immunocomplexes were visualized with an Odyssey Infrared Imaging System (LI-COR).

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, & Kim H. (2017). The transcription factor MafB promotes anti-inflammatory M2 polarization and cholesterol efflux in macrophages. Scientific Reports, 7, 7591.

Publication 2017

anti-β-actin anti-GAPDH (sc-25778; anti-Stat6 (sc-621; anti-pStat6 anti-LXRα anti-Flag (M2) or anti-HA Affinity Gel goat anti-rabbit IgG-HRP IRDye 800CW–conjugated goat anti–mouse and anti–rabbit IgG IRDye 800–conjugated anti-Flag and anti-HA Odyssey Infrared Imaging System

Actin Anti igg Antibodies Cells Chip Chip assays Gapdh Goat Immunoblot Irdye 800 Irdye 800cw Mafb Mouse Primers Rabbit Rt pcr Stat6 Tissues Trizol

Corresponding Organization : Massachusetts General Hospital

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Variable analysis

independent variables

Flag-tagged MafB purified with M2 Affinity Gel
Myc/DDK-tagged LXRα obtained from OriGene

dependent variables

RT-PCR gene expression
ChIP assay results

control variables

Cells and tissues processed and analyzed as previously described
Total RNA extracted using TRIzol method
Primer sequences for RT-PCR and ChIP assays listed in Supplementary Tables S2 and S3
Antibodies used for immunoblot and ChIP analyses: anti-β-actin, anti-GAPDH, anti-MafB, anti-Stat6, anti-pStat6, anti-LXRα, anti-Flag, anti-HA

controls

Positive control: No information provided
Negative control: No information provided

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