The cDNA amplicons from total RNA extracted from a mouse brain cortex were inserted in a TOPO-pCR4 vector (Invitrogen, Carlsbad, CA, USA) followed by linearization and in vitro transcription in presence of dUTP-digoxigenin (Roche, Basel, Switzerland) according to the manufacturer instructions (Invitrogen, Carlsbad, CA, USA).
Relative standard curves were produced by ten-fold serial dilutions of liver RNA extracted from a mouse receiving PBS.
Total protein extract from a mouse E16.5 embryo was collected after homogenization in 1 mL of tissue lysis buffer (0,25% Triton X-100, Complete (Roche Diagnostics) in PBS), sonication, and centrifugation.
Matrigel is a commercial product extracted from a mouse sarcoma rich in extracellular matrix proteins.
One of the first commercially available hydrogels onto the market was Matrigel™ (BD Biosciences), which is based on an ECM extracted from a mouse sarcoma (Kleinman et al. 1986).
Thus, the scattering coefficient was almost three times higher than that typically measured immediately after the brain is extracted from a mouse and is post-fixed.
Direct evidence for the effect of miR-34c on cyclin E regulation was provided by using cells extracted from a mouse model overexpressing cyclin E, who develops multiple pulmonary adenocarcinomas, and human LC cell lines transfected with miR-34c resulting in repression of cell proliferation [ 77].
Briefly, a reference design was used for microarray hybridization: intestinal and liver RNA extracts from all mice were pooled in an equimolar proportion and used as the reference sample.
Furthermore, we detected NF-κB binding in colonic nuclear extracts from all but one of the CONV-R mice we assessed, but never in any extract from a GF mouse.
Total RNA was extracted from all mouse livers using an RNeasy Mini kit and DNeasy Tissue kit (QIAGEN Inc., Valencia, CA, USA).
