IFNγ enhances macrophage host cell death after type I (RH) and type II (Pru) Toxoplasma gondii (Tg) infection. Graphs show cell death as measured by LDH release assay (left) and viability measured by XTT assay (right) in untreated or IFNγ‐primed THP‐1 cells infected with type I or type II Tg for 24 h.
LDH release assays from THP‐1 cells left untreated or primed with IFNγ, transfected with siRNA against indicated GBPs or non‐targeting control (CTRL), and infected with indicated strains of Tg for 24 h.
LDH release assay from primary monocyte‐derived macrophages (MDM) left untreated or treated with IFNγ, transfected with siRNA against GBP1 or non‐targeting control (CTRL), and infected with indicated strain of Tg for 24 h. Mean ± SEM of n = 4 donors shown.
LDH release assay from indicated wild‐type, GBP1‐knockout (ΔGBP1), or GBP1‐reconstituted (ΔGBP1 + Tet‐GBP1) cells infected with type I or type II Tg for 24 h. Cells were untreated or treated with IFNγ or additionally treated with Doxycycline (Dox) as indicated.
Real‐time propidium iodide (PI) uptake assay from the indicated THP‐1 cells infected with type I or type II Tg, and untreated or treated with IFNγ and/or Dox as labeled. AU, arbitrary units.
LDH release assay from wild‐type THP‐1 or ΔGBP1 cells stably reconstituted with Dox‐inducible expression plasmids of the indicated mutants of GBP1. Cells were pre‐treated with IFNγ and Dox and infected with either type I or type II Tg for 24 h.
Data information: Graphs in (A, B and D‐F) show mean ± SEM from
n = 3 independent experiments.
*P ≤ 0.05;
**P ≤ 0.01
, ***P ≤ 0.001,
****P ≤ 0.0001 for indicated comparisons in (A–D) from two‐way ANOVA or comparison between THP‐1 WT and other genotypes by one‐way ANOVA in (F) following adjustment for multiple comparisons; ns, not significant.
