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Multidimensional pooled shRNA screens in human THP-1 cells identify candidate modulators of macrophage polarization

Fig 1

THP-1 monocyte to macrophage differentiation and M1 / M2 polarization.

(A) Schematic representation of treatments to differentiate THP-1 monocytes (Mo) into macrophages (M0) or polarized macrophages (M1, M2). (B) Morphological and cell surface marker expression (CD11c, CD11b) changes observed by microscopic and flow cytometric analysis of PMA (phorbol 12-myrisate 13-acetate) differentiated M0. FCS: forward-scatter; SSC: side-scatter. (C) Resting CD38- / CD209- M0 macrophages (top panel) are polarized in CD38+ / CD209- M1 fraction upon LPS / IFNγ treatment (middle panel) and in CD38- / CD209+ M2 fraction upon IL4 / IL13 treatment (bottom panel). (D) Cytokine and chemokine release profiling of THP-1 M1 and M2 macrophages detecting M1 (black) and M2 (grey) specific soluble mediators, respectively. Supernatants were collected from three independent experiments and then pooled for the analysis. *Exogenously added to the M2 polarization medium. (E) Correlation analysis of transcriptional fold change (FC) during Mo to M1 (top panel) or M2 (bottom panel) polarization comparing THP-1 and human primary cells. For macrophage polarization MCSF (human primary cells) or PMA (THP-1) differentiated macrophages were treated with IFNγ (M1) or IL4 (M2). Whole genome microarray measurements were used from three independent experiments and each dot corresponds to a probe on the array. Highlighted genes are previously reported human markers relevant for the M1 or M2 polarization [32,31] (additional references listed in the main text). Highlighting was limited to genes which at least in one of the cellular models had at least one probe with 1 < or– 1 > FC. Dashed line is the y = x reference, solid line is fitted on the experimental data y = a + b * x (a = interception of y, b = slope).

Fig 1

doi: https://doi.org/10.1371/journal.pone.0183679.g001