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. 2017 Oct 24;18(12):2172–2185. doi: 10.15252/embr.201744154

Figure 6. BCKAs are taken up by human monocyte‐derived macrophages and reduce phagocytosis.

Figure 6

  • A–C
    Leucine (A), valine (B), and succinate (C) labeling patterns were determined by GC‐MS in cell extracts from M‐CSF differentiated macrophages cultured with 100 or 300 μM of 13C‐αKIC and 13C‐αKIV. n = 3 technical replicates. Monocytes were isolated from two independent donors (d1, d2). M0, M1 to Mn: M is the base mass of an ion fragment, and the following number from 0 to n (active carbon number) indicates the mass shift from M.
  • D, E
    Monocyte‐derived macrophages previously incubated in the absence (medium control) or presence of 50, 100, 200, or 300 μM of BCKAs (KIV, KIC, and KMV) at 37°C for 24 h were incubated with 1‐μm‐diameter fluorescent beads at 37°C for 2 h, and phagocytosis of beads was examined by fluorescence microscopy. (D) The number of beads engulfed by each cell is quantified using ImageJ software. Line represents mean. Cells were isolated from two independent donors, and eight fields were examined per condition (n > 165 cells). Mann–Whitney test. (E) Fluorescent beads (green). Plasma membrane (red) is stained with Cell Mask stain. The Hoechst stains nuclei (blue). Scale bar: 40 μm.