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. 2019 Oct 4;10:2210. doi: 10.3389/fimmu.2019.02210

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Copyright © 2019 Herdoiza Padilla, Crauwels, Bergner, Wiederspohn, Förstner, Rinas, Ruf, Kleemann, Handrick, Tuckermann, Otte, Walther and Riedel.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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PMA-activated THP-1 cells allow investigation of phagocytosis of opsonized latex beads by human MΦ. (A) Schematic representation of the workflow for differentiation, transfection, and analysis of phagocytosis of FITC-labeled, opsonized latex beads for high-content screening of a miRNA library and subsequent experiments. (B–D) Bright-field microscopy and FACS analysis of untreated (Mo) or PMA-activated (MΦ) THP-1 cells. (B) Microscopic images were acquired with an AxioObserver Z1 microscope (Zeiss) using a 40× objective. Scale bars represent 20 μm. (C) Histogram plots of one representative experiment and (D) quantification of the percentage of FITC⁺ (left) and mean fluorescence intensity (MFI) of FITC⁺ cells (right) of n = 6 independent experiments. In (D), values from each experiment are indicated by symbols and the mean ± standard deviation (SD) are indicated by a bar and whiskers. Statistical analysis was performed using Student's t-test (***p < 0.001).