Supplementary Figure 3: Analysis of lower motor neuron pathology and thalamic lesions in Cx3cr1CreER/+Tgfbr2fl/fl→DTA chimeras.
(a) Analysis of innervation of neuromuscular junctions in the tibialis anterior muscle of Tgfbr2fl/fl→DTA and Cx3cr1CreER/+Tgfbr2fl/fl→DTA chimeras. Innervation was quantified by assessing overlay of synaptic vesicle protein (SV2) and neurofilament (NF) 165 kDA staining with α-BTX labeled endplates. Positive control is tibials anterior muscle from SODG93A mice (day 140) illustrating complete denervation. Lines in graph are mean values of n = 4, 2, 3, 4 mice/group and at least 40 motor endplates/mouse. Scale bar 50µm. (b) Enumeration of cresyl violet stained motor neurons (>200μm2) in the dorsal horns of the cervical enlargement. Lines in graph are mean values of n=4 mice/group and at least 5 sections/mouse. NS=not significant P=0.1633 Student’s two-tailed unpaired t-test. Scale bar 100µm. (c) Enumeration of VAChT+ motor neurons in the ventral horns of the spinal cord. Representative images are taken from the lumbar enlargement. Lines are mean values of n= 4, 3, 4 mice and at least 4 sections/mouse. * P= 0.0354 (cervical) and * P=0.0487 ** P=0.0034 (lumbar) 1-way ANOVA Dunnett’s Multiple Comparison test. Scale bar 200μm. (d) Hematoxylin and eosin staining visualizing lesions in the thalamus in end stage KO→DTA mice. Lesions were quantified based on the presence of giant cells (white arrow) with an area >150µm2. Lines are mean values of n=3 mice/group and 2 sections/mouse. Scale bar 100µm (top panel) and 25µm (bottom panel). (e) F4/80 and fluoromyelin staining in the thalamus. Images are representative of n=3 mice/group. Scale bar 500µm and 50µm. (f) Confocal images of F4/80 and fluoromyelin staining in the thalamus. Images are representative of n=3 mice/group. Scale bar 10µm.
