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. 2016 Oct 12;11(10):e0164544. doi: 10.1371/journal.pone.0164544

Fig 3. Tomosyn is expressed in the rat hypothalamo-posterior pituitary axis.

Fig 3

(a) Presence of tomosyn in the rat anterior and posterior pituitary as indicated by western blotting. Pituitary lysates were separated on SDS-PAGE followed by western blotting using anti-tomosyn, anti-SNAP25, anti-syntaxin 1A, anti-syntaxin 1B, anti-AVP, and anti-β-actin antibodies (as a loading control). (b) Subcellular fractionation of rat posterior pituitary samples. Fraction 1 (cytosolic proteins, F1) and Fraction 2 (membranes and membrane organelles, F2) were subjected to SDS-PAGE followed by western blotting using anti-tomosyn antibody. Five μg of proteins was loaded onto each lane. The subcellular fractionation was confirmed by western blotting with anti-Akt (cytosolic marker) and anti-N-cadherin (membrane fraction marker) antibodies. Tomosyn was present in both fractions. (c) Expression levels of tomosyn-1 mRNA in rat cortex and hypothalamus are similar. The amount of mRNA was determined using quantitative RT-PCR. The values are normalized to β-actin mRNA and are expressed as the mean ± SEM.