Computational protocol: Bim and Bmf Synergize To Induce Apoptosis in Neisseria Gonorrhoeae Infection

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Protocol publication

[…] Cells seeded on coverslips were infected for 15 h, fixed in 3.7% PFA and permeabilized using 0.1% Triton X-100. Nonspecific binding was blocked by using 1% goat serum. The samples were stained using anti-actin (Sigma) and anti-tubulin (molecular probes) followed by detection with fluorochrome-coupled secondary antibodies (Jackson Immuno Research) using a Leica confocal microscope with TCS software or a Zeiss immunofluorescence microscope with ACT software. 3dimensional remodeling was performed using Metamorph and Imaris software.For apoptosis quantification, fixed cells were stained with 1 µg/ml Hoechst 33342 (Invitrogen) for 10 min followed by intense washing with PBS. A minimum of 5 fields per slide was analyzed for chromatin condensation using a Zeiss immunofluorescence microscope. […]

Pipeline specifications

Software tools MetaMorph, Imaris
Application Laser scanning microscopy
Organisms Neisseria gonorrhoeae
Diseases Infection