Anti CTL1 antibody could detect the full-length (200-k Da) recombinant L1 in the lysate of transfected High5 insect cells (lane 3), whereas no positive band was detectable in the mock-transfected (control) High5 lysate (lane 4). Omission of primary antibody (-anti CTL1) resulted in no immunostaining (substantia nigra). Blocking of anti CTL1 antibody with epidermal growth factor (EGF) did not interfere with the immunostaining in the neighboring section of (b). Absorption of the antibody with L1 gene-transfected membrane (L1m) completely eliminated the immunostaining, whereas absorption of the antibody with mock-transfected membrane (Mock) had no effect (d'-f').
d,d': stria terminalis; e,e': substantia nigra; f,f': cerebellar cortex. The two polyclonal antisera (anti CTL1 and anti FLL1 antibodies) demonstrated an analogous staining pattern on immunohistochemistry, though the latter antiserum tended to stain passing proximal axons better than the former (Fig. Consequently, described here are the results obtained with the anti CTL1 antibody unless specified otherwise.
e and f, Bright-field photomicrograph showing L1 neurons in the lateral posterior thalamic nucleus of the upper brainstem.
a,a',b,b': Bar = 25 μm; c,c',d,d',e: Bar = 200 μm; f: Bar = 50 μm. The immunoreactivity of fibres was generally strong and this type of staining was observed chiefly in the major fibre bundles.
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For this reason, we studied the total distribution of L1 in the adult mouse CNS using specific polyclonal antisera against full-length L1 and the C-terminal cytoplasmic domain of L1 at the light microscopic level.The accumulation of immunoreactive L1 was greatest in a non-neuronal element of the major fibre bundles, the lateral olfactory tract, olfactory and temporal limb of the anterior commissure, corpus callosum, stria terminalis, globus pallidus, fornix, mammillothalamic tract, solitary tract, and spinal tract of the trigeminal nerve.High to highest levels of non-neuronal and neuronal L1 were found in the grey matter; the piriform and entorhinal cortices, hypothalamus, reticular part of the substantia nigra, periaqueductal grey, trigeminal spinal nucleus etc.It is the world’s first gay monument which stands as a reminder of gay persecutions and discrimination due to their sexual orientation and to support the fight of the LGBT community for equality.The memorial with three pink triangles was built in 1987 to commemorate gay and lesbian victims of the Nazi regime but the idea was inspired by the arrest of gay and lesbian activists for trying to place a wreath at the National War Memorial on the Dam Square in 1970.