Fig 1: Overexpression of IGFBP-rP1 affects the polarization of EC-related macrophages by regulating the PI3K/AKT pathway. Human THP-1 monocytes were treated with PMA for 24 h to induce M0 macrophages. Then, M0 cells were co-cultured with EC cells overexpressing IGFBP-rP1 and/or overexpressing AKT for 48 h to produce TAMs. The level of M1 markers, (A) iNOS and (B) TNF-α and M2 markers, (C) CD163, (D) Arg-1 and (E) MR were analyzed using ELISA. *P<0.05. IGFBP-rP1, insulin-like growth factor binding protein-related protein 1; EC, endometrial carcinoma; TAMs, tumor-associated macrophages; iNOS, inducible nitric oxide synthase; NC, negative control; Arg-1, arginase-1; MR, mannose receptor.
Fig 2: Correlations between IL-10, RANTES, CD206 CD163, FAP, Arginase 1, and TGF-β1 with the soluble immune checkpoint molecules.
Fig 3: Clinical relevance of sTREM2 in childhood IgAN. (a,b) Comparison of plasma sTREM2 and sCD163 level in IgAN (n = 38) versus healthy individuals (n = 18). (c) Correlation analysis of plasma sTREM2 and sCD163 (n = 38). (d,e) Correlation of sTREM2 and sCD163 with the amount of 24 h proteinuria (n = 38). (f,g) Comparision of plasma sTREM2 and sCD163 abundance among IgAN patients with different pathological grading. Data were presented as mean values + SD. Abbreviations: IgAN, IgA nephropathy; sCD163, soluble CD163; sTREM2, soluble TREM2.
Fig 4: Significant anti-inflammatory TAMs are detected in triptolide-treated tumors that receive IL-4.The tumor was dissected out and dissociated into single cells at sacrifice. Afterwards the single cell preparation underwent FAC analysis, showing significant increases in pro-inflammatory fraction (CD163-) of the macrophages (F4/80+) in the tumors from Triptolide-treated mice, which were abolished by addition of IL-4 treatment.
Fig 5: Triptolide reprograms macrophages towards a pro-inflammatory status.We used the concentration of Triptolide (TP; 55nM) that was found to induce IC50 of ovarian cells to treat naïve macrophages, or naïve macrophages that had been polarized to M2 by IL-4. (A–B) CCK-8 assay for naïve macrophages (naïve M ϕ ; A) and for IL-4-primed naïve macrophages (IL-4-M ϕ ; B). (C–D) Cell clock assay to analyze naïve M ϕ and IL-4-M ϕ treated by 55nM Triptolide, shown by quantification (C), and by representative images (D). (E–F) Naïve M ϕ and IL-4-M ϕ were treated by 55nM Triptolide. TUNEL staining was done to assess apoptosis, shown by representative images (E) and by quantification (F). (G) Flow cytometry analysis of macrophage polarization by CD163 and CD86. (H) ELISA for IL-1 β , TNF α , IFN γ , CD163, IL-10 and Nrf2. ⁎p < 0.05. ns: non-significant. N = 5. Scale bars are 20 μ m.
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