Fig. 6.

The effect of heat stress on the production of pro-inflammatory cytokines, ROS accumulation and apoptosis. The cells were infected with B. abortus and subjected to heat exposure. The production of pro-inflammatory cytokines and ROS was evaluated at 24 h pi while the apoptosis assay was performed at 48 h pi. Levels of pro-inflammatory cytokines: Tnf, Il6, Il10, Il1b and Mcp1 in RAW 264.7 (a) and BMM (b) through RT-PCR. Production of ROS after heat exposure was measured by fluorescence microscopy (c) and spectrometry (d). The infected cells were treated with inhibitors of mitochondrial respiratory chain (TTFA) or NADPH oxidase (DI) (e). The apoptosis was evaluated by activation of caspase-3 (f) and flow cytometry (g) after heat exposure. The intensity of FITC was also evaluated from random 10,000 cells (h). Data represent the mean ± SD of triplicate experiments. Asterisks indicate significant differences (p < 0.01). (Scale bar = 20 μm)