Hence, we characterized the cytokine profile during K. rhinoscleromatis infection by measuring the production of some of them in mice lung extracts.
The nucleic acid demethylase activity of the mouse lung nuclear extracts was assayed as their ability to demethylate 3-methylcytosine in [14C]-methylated poly dC).
Moreover, the in vitro nucleic acid demethylase activity of mouse lung nuclear extracts was increased by harman and norharman, two known inhibitors of MAO present in tobacco smoke [24], [25].
Mouse lungs were extracted, shock-frozen in liquid nitrogen, and kept at −80°C until processing or fixed by intratracheal instillation of 4% paraformaldehyde in PBS and embedded into paraffin for hematoxylin-eosin (H&E) staining.
Briefly, viral RNA of mouse lung homogenate was extracted using the RNeasy Mini Kit (Qiagen, Valencia, CA, USA).
To quantify Mp rapidly from infected tissues, we developed a real-time PCR-based method for measuring the numbers of Mp in extracts of mouse lung.
Membrane proteins extracted from biofilm or planktonic grown cells conferred protection in the mouse lung, while only the biofilm extract induced rBipA antibodies, albeit at lower levels compared to vaccination with rBipA.
Total RNA was extracted from mouse lung and from mouse embryonic day 13 fibroblasts using TRIzol (Invitrogen Corp).
Lipids to be identified were extracted from mouse lung tissue by homogenization of the tissue in 70% ethanol.
