In addition, confocal microscopy showed co-localization of the pre-synaptic marker synapsin and γ-secretase active sites in mouse embryonic cortical neurons.
We are also very thankful for the technical assistance from Dr. Kjell Hultenby (electron microscopy) and Birgitta Wiehager (preparation of mouse embryonic cortical neurons).
Rat and mouse embryonic cortical neurons were prepared using the papain disassociation technique described previously [55] and cultured on 8-well, coverglass-bottom dishes for 3 7 days prior to analyses.
Primary cultures of mouse embryonic cortical neurons were prepared as described previously.
Cultured primary mouse embryonic spinal motor neurons were transduced with recombinant lentivirus, cultured for 5 days and fixed and counted.
This concept has been recently revisited with the detection of large amounts of 5-hydroxymethylcytosine, now considered as the sixth DNA base, in mouse embryonic stem cells, Purkinje neurons and brain tissues.
Therefore, to investigate the potential role of retinoid signalling in Aβ production, we treated mouse embryonic day 15 cortical neurons with dexamethasone and retinoid agonists (Fig. 1).
Here, we identified candidate target genes bound and regulated in mouse embryonic mDA progenitors and neurons at E12.5, which we will refer to as the in vivo data set, as distinct from the published in vitro data set.
Live cell imaging experiments revealed that in mouse embryonic fibroblasts (MEF) and neurons growing in primary midbrain cultures isolated from WT, Nrf2-KO, Keap1-KO or Keap1-KD mice [ 41– 44], disruption of Nrf2 results in a significant loss of the basal ΔΨm [ 23].
Indeed, several in vitro studies, using a wide variety of cell lines, such as Chinese hamster ovary (CHO) cells, primary neurons, mouse embryonic fibroblasts (MEFs), HeLa cells (Zhang et al., 2009), HC11 mammary epithelial cells and HEK293T cells demonstrated that BOK overexpression could induce apoptosis.
