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. 2022 Mar 2;12(3):210339. doi: 10.1098/rsob.210339

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© 2022 The Authors.

Published by the Royal Society under the terms of the Creative Commons Attribution License http://creativecommons.org/licenses/by/4.0/, which permits unrestricted use, provided the original author and source are credited.

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Figure 3. — DA neurons express the calcium-sensors Syt1, Syt4 and Syt7. (a) Protocol used for double immunostaining for two primary antibodies from the same host (adapted from a Jackson ImmunoResearch protocol). Use of normal rabbit serum and unconjugated Fab fragments for blocking after the first secondary. (b–d) IHC of midbrain and striatal slices of adult DAT^IREScre heterozygote mice showing colocalization of VMAT2 and either Syt1, Syt4 or Syt7 in DAergic neurons. Scale bar = 20 μm. (e) Control images were obtained using the full protocol without the use of the second primary antibody (in the midbrain).