Fig. 1
High-content imaging with IncuCyte to assess intracellular Mtb growth in human host cells. (A) The role of histone acetylation in transcriptional regulation of T cells (left) and Mtb-infected macrophages (right). Virulent Mtb typically induces deacetylation, chromatin compaction and inhibition of gene expression. Histone acetyltransferases (HAT), HDAC inhibitors (HDACi), acetyl group (Ac), transcription factors (TF). (B) Five classes containing 18 different HDAC proteins and their potential effects on cellular functions and immune cell responses. (C) Schematic illustration of the macrophage infection model, mycobacterial strains and experimental setup as well as data acquisition and analyses. (D) Longitudinal response of the Mtb infection control (MOI1), antibiotics control (RIF + INH) and internal HDAC inhibitor (PBA) control. Intracellular growth of H37Rv-GFP in hMDMs (Total integrated intensity expressed as GCU x µm2/image) was monitored in real-time (day 0–5) using IncuCyte (median +/- error). (E) The assay Z’ factor was determined using the mean +/- standard deviation of the negative (MOI1 infection) and positive (RIF + INH) controls. Representative data from one donor out of n = 3 is shown. (F) Viability of uninfected or H37Rv-infected hMDMs treated with RIF + INH or PBA. Data from n = 6 donors are presented in the bar graph (median +/- range). Host cell viability was monitored with Cytotox-red. (G) Representative microscopy images illustrating hMDMs (grey color) infected with H37Rv-GFP (green color) and treated with RIF + INH or PBA. Magnification x20. Illustrations in (A-B) were created with Biorender.com.
