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. 2015 Nov 1;13(9):515–528. doi: 10.1089/adt.2015.664

Fig. 1.

Fig. 1.

Imaging dual toll-like receptor (TLR) and bacterial infection-induced mitogen-activated protein kinase (MAPK) response in mouse macrophages. Wild-type (WT) immortalized murine macrophages (IMMs) were either (A) stimulated with dual TLR ligands—10 nM each of P3C and lipid A—or (B, C) infected with live or formalin-killed (FK) Burkholderia cenocepacia J2315 at multiplicity of infection (MOI) 10 for up to 4 h and the ratio of nuclear to cytoplasmic intensity of the MAPK-activated transcription factor, phosphorylated activating transcription factor 2 (pATF2), was calculated. (D) The median nuclear-to-cytoplasmic ratios of ATF2 normalized to maximum. (E, F) Images at 20× magnification captured by the CellInsight NXT are shown for untreated and dual ligand treatment for 60 min. Data shown are representative of two independent experiments (D; median+median absolute deviation), and number of cells imaged were 1,964 cells for dual TLR ligand treatment, 2,106 cells for live bacterial infection, and 2,009 cells for killed bacterial infection.