Journal: FEMS Immunology & Medical Microbiology

Article Title: Chlamydia (Chlamydophila) pneumoniae-induced cell death in human coronary artery endothelial cells is caspase-independent and accompanied by subcellular translocations of Bax and apoptosis-inducing factor

doi: 10.1111/j.1574-695x.2006.00083.x

Figure Lengend Snippet: Fig. 4. Subcellular translocation of Bax during Chlamydia pneumoniae infection of human coronary artery endothelial cells. (a) Cells were infected with Chamydia pneumoniae T45 at multiplicities of infection 1.25 or 10 for 8 h and incubated with N-20 polyclonal antibody towards Bax and secondary antibody. Cells with a punctate fluorescence pattern were enumerated by fluorescence microscopy. Bars represent mean standard deviation of six or seven experiments. P o 0.05. (b) Subcellular localization of Bax in uninfected cells showing diffuse cytosolic fluores- cence. (c) Subcellular localization of Bax in cells infected with C. pneumoniae, multiplicity of infection 1.25, for 24 h showing punctate cytosolic fluorescence.

Article Snippet: Cells were washed twice in phosphate-buffered saline (PBS) and fixed in 4% paraformaldehyde in PBS for 15 min. After washing, cells were incubated for 30 min in N-20 antiBax rabbit polyclonal antibody (Santa Cruz Biotechnology, Santa Cruz, CA) at 1 : 50 dilution in permeabilization buffer (bovine serum albumin (BSA) 1 g L 1, 0.05% saponin in PBS), washed twice in PBS-BSA, blocked in 4% normal goat serum for 30 min, washed in PBS-BSA and visualized by incubation for 30 min with a 1 : 100 dilution of Alexa Fluor 568 F(ab0)2 goat antirabbit IgG antibody (Molecular Probes, Eugene, OR).

Techniques: Translocation Assay, Infection, Incubation, Microscopy, Standard Deviation

Journal: FEMS Immunology & Medical Microbiology

Article Title: Chlamydia (Chlamydophila) pneumoniae-induced cell death in human coronary artery endothelial cells is caspase-independent and accompanied by subcellular translocations of Bax and apoptosis-inducing factor

doi: 10.1111/j.1574-695x.2006.00083.x

Figure Lengend Snippet: Fig. 5. Subcellular translocation of apoptosis-inducing factor (AIF) dur- ing Chlamydia pneumoniae infection of human coronary artery endo- thelial cells. (a) Cells were infected with Chlamydia pneumoniae T45 at multiplicities of infection 1.25 or 10 for 8 h and incubated with N-19 polyclonal antibody towards AIF and secondary antibody. Cells with nuclear fluorescence were enumerated by fluorescence microscopy. Bars represent mean standard deviation of nine to 11 experiments. P o 0.05. (b) Subcellular localization of AIF in uninfected cells showing cytosolic fluorescence. (c) Subcellular localization of AIF in cells infected with C. pneumoniae, multiplicity of infection 1.25, for 24 h showing nuclear fluorescence.

Article Snippet: Cells were washed twice in phosphate-buffered saline (PBS) and fixed in 4% paraformaldehyde in PBS for 15 min. After washing, cells were incubated for 30 min in N-20 antiBax rabbit polyclonal antibody (Santa Cruz Biotechnology, Santa Cruz, CA) at 1 : 50 dilution in permeabilization buffer (bovine serum albumin (BSA) 1 g L 1, 0.05% saponin in PBS), washed twice in PBS-BSA, blocked in 4% normal goat serum for 30 min, washed in PBS-BSA and visualized by incubation for 30 min with a 1 : 100 dilution of Alexa Fluor 568 F(ab0)2 goat antirabbit IgG antibody (Molecular Probes, Eugene, OR).

Techniques: Translocation Assay, Infection, Incubation, Microscopy, Standard Deviation