In 2011, Matthew Wood and colleagues at the University of Oxford used both approaches to show that they could use exosomes to downregulate neuronal gene expression in the mouse brain by loading the exosomes with a neuron-targeting peptide and specific short interfering RNAs.
Moreover, Wnt target gene expression induced by loading in WT mice was absent in TG mice.
In contrast to WT mice, osteopontin expression levels were only down-regulated by loading in tibiae from WT-OVX mice and remained unchanged in tibiae from WT-SN and ERα−/− mice at all time points after loading.
Bone marrow and spleen suspension from C57BL/6J mice were attached to microscope slides by loading 200 μl of cell suspension into cytofunnels (Thermo Electron Corporation) and centrifugation in a Shandon Cytospin 4 Cytocentrifuge (Thermo Scientific) for 10 minutes at 800 rpm.
WT responder T cells were prepared from pooled spleen and LN cells from CD90.1 congenic mice by MACS, and were loaded with CFSE.
Consistent with this finding the transcription of mammalian core histone gene, H4, was up-regulated by loading in WT mouse tibia.
We demonstrate here that strain-related Sost down-regulation in cells of the human osteoblastic Saos2 cell line recapitulates in vitro, in terms of time, that stimulated in osteocytes by loading of the mouse tibia in vivo [9].
In tibiae of ERα−/− mice, the number of genes differentially regulated by loading was markedly reduced at all time points.
The difference in drug activity between the two mouse strains was even more pronounced with MXF treatment at 100 mg/kg, which reduced the bacterial load in C57BL/6 mice by 0.85 log10CFU and by more than 3.5 log10CFU in GKO mice after 7 days of treatment when compared to untreated mice at the start of treatment (P<0.05).
We compared expression of angiogenic and survival protein in tumor lysates from control and treated mice by western blotting analysis using β-actin as loading control (Fig. 4).
