Journal: Cells

Article Title: Statins Modulate Microenvironmental Cues Driving Macrophage Polarization in Simulated Periodontal Inflammation

doi: 10.3390/cells12151961

Figure Lengend Snippet: MyD88−dependent modulation of M2 polarization by simvastatin. ( A ) A schematic representation of simvastatin, MyD88 inhibitor treatment, and THP-1-macrophages (M0–M1–M2). ( B ) Cell surface antigen analysis of M2 macrophage markers CD68, CD163, and CD206 was conducted using flow cytometry. Expression levels were normalized using IgG isotype control, and values were expressed in percentage of positive cells (mean ± SD) from three independent preparations. ( C ) RT-PCR analysis and heat maps summarizing TLR and NFkB signaling from M0 (CCL2), M1 (LPS, IFNγ, TNFα), and M2 (IL4, IL10, TGFβ) macrophages. (n = 3) preparations per treatment group, and data found to be significantly different were plotted as heat maps. ( D ) Graphical summary of statin-mediated suppression of TLR and NFkB signaling.

Article Snippet: Phenotyping of macrophages was performed in 1% BSA and 3% human serum PBS according to standard methods using a panel of antibodies targeting CD68 (R and D Systems Cat# IC20401P, Minneapolis, MN, USA, RRID: http://scicrunch.org/resolver/AB_2074835 , accessed on 28 April 2023), CD163 (R and D Systems Cat# FAB1607P, RRID: http://scicrunch.org/resolver/AB_2074536 , accessed on 28 April 2023), and CD206 (R and D Systems Cat# FAB25342P, RRID: http://scicrunch.org/resolver/AB_10889015 , accessed on 28 April 2023); antibodies all from R and D Systems.

Techniques: Flow Cytometry, Expressing, Control, Reverse Transcription Polymerase Chain Reaction

Journal: International Journal of Molecular Sciences

Article Title: Neuropeptide Y Promotes Human M2 Macrophage Polarization and Enhances p62/SQSTM1-Dependent Autophagy and NRF2 Activation

doi: 10.3390/ijms232113009

Figure Lengend Snippet: Flow cytometric analysis of the M1 markers HLA-DR and CD16 and the M2 markers CD206 and CD163 on ( a ) M0, ( b ) polarized M(IFN-γ/LPS), and ( c ) polarized M(IL-10) macrophages. Macrophages (7 × 10 5 cells per mL) were stimulated or not with 10 −8 M NPY in the presence or absence of LPS in complete medium for 24 h and then analyzed for surface marker expressions by flow cytometry. Histograms show the percentages of positive cells (%) and the mean fluorescence intensity (MFI). Results are expressed as mean value ± SD of 4 independent experiments. Significance was determined by one-way ANOVA followed by Tukey’s post hoc analysis; *: p < 0.05, **: p < 0.01.

Article Snippet: To determine macrophage phenotypic surface markers, macrophages were stained with the following monoclonal antibodies (mAbs): phycoerythrin (PE)-CD163, fluorescein isothiocyanate (FITC)-CD206 and PE-Vio770-CD206 mAbs (Miltenyi Biotec), allophycocyanin (APC)-CD16 and APC-Alexa Fluor 750-HLA-DR (clone Immu357) mAbs (Beckman Coulter, Lane Cove, Australia), PE-CD1a and FITC-CD14 mAbs, or with isotype-matched control mAbs from PharMingen (PharMingen, San Diego, CA, USA) for 30 min at 4 °C.

Techniques: Marker, Flow Cytometry, Fluorescence