Efficient co-expression of secretory BMPs, Noggin, and Venus in the cortex and the hippocampus of rAAV injected mice at DPI 14. A. Confocal fluorescent immune histological images of sections from rAAV infected adult mouse brains (hippocampus and cortex) demonstrated cytosolic expression of Venus (green) and v-BMPs or v-Noggin (red) visualized by anti-GFP and anti-2A antibodies, respectively. The insets in the middle row show the expression of v-BMPs or v-Noggin in glia and neurons in the stratum radiatum (St.r) at high magnifications. In the insets of the bottom row, overlay of the CA2 region in the hippocampus is depicted demonstrating the nuclear exclusion of the 2A-tagged peptide factors (yellow) in cells expressing Venus (green) in the nucleus and the soma. A. Scale bar, 500 μm. Scale bar inserts, 15 μm. B. Fluorescent immune-histological images show the nuclear excluded 2A-tagged v-BMP7 and v-BMP2 in rAAV infected neurons of cortical layer V/VI. Scale bar, 20 μm. C. Confocal fluorescent immune-histological images of sections from rAAV infected adult mouse brains show expression of v-Venus and membrane associated v-sVenus in the hippocampus and cortex. Lower panel: High magnifications of the hippocampal CA1 region of the somatic and nuclear excluded Venus immunoreactivity. Scale bar, 500 μm. Scale bar insets, 15 μm. D. After rAAV infection immunoblots detected precursors and mature protein factors in hippocampal extracts. Precursors (Prec.), dimerized Noggin (dNoggin) and mature proteins are indicated by arrowheads. Beta-actin was used as loading control.
Heinonen et al. BMC Neuroscience 2014 15:38 doi:10.1186/1471-2202-15-38