A) RT-PCR results show presence of the full-length (281 bp) and truncated (353 bp) TrkB
isoforms in preparations from brain (lane 1), cerebellum (lane 2), and isolated mouse
glomeruli (lane 3 and 4). MWM: molecular weight marker; lane 5: negative control, i.e. PCR
products obtained from the osteoblast cell line MC3T3-E1; lane 7: PCR negative
control.
B) Western Blot (WB) analysis was conducted on protein extracts from mouse brain (1), cerebellum (2)
and two preparations of isolated glomeruli (3, 4) in non-reducing conditions. A band slightly higher
than 120kDa, corresponding to the full length molecule, is present in all lanes. In
addition, higher bands are observable. Bands of lower molecular weight can be detected as
well. MWM = molecular weight marker, Lane 5 = negative control, performed by loading
protein lysates from the osteoblast cell line.
C) Immunogold, conducted on a cryosection from control mouse kidney, shows TrkB
positivity in a podocyte (red circles) and a dot in an endothelial cell (light blue
circle). Scale bar 500nm.
D) Double staining shows that TrkB mostly co-localizes with nephrin (upper panels) and
seldom with CD31 (lower panels, arrow). Scale bars 10μm.