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. 2019 Jul 29;6(4):ENEURO.0237-19.2019. doi: 10.1523/ENEURO.0237-19.2019

Figure 2.

Figure 2.

Morphine upregulates FHC protein in the neuronal cytoplasm. A, FHC is expressed in the soma and processes of morphine-treated neurons. Neurobasal cultures were treated with morphine (1 µM, 24 h) or vehicle before fixation and immunostaining. Cultures were immunostained for FHC (green) and the neuronal marker β-III Tubulin (red), and counterstained with the nuclear marker Hoechst (blue). Images were acquired with 20× and 60× objectives. Morphine treatment visibly increased FHC staining in the soma and processes. One group of neurons was immunostained without both primary antibodies, showing that non-specific staining was negligible. B, Morphine upregulates FHC in cytoplasmic extracts of neurobasal cultures. Cultures were treated with morphine (1 µM, 3, 6, or 24 h) or vehicle, and separated into cytosolic and nuclear extracts. Morphine dose dependently increased FHC protein levels in cytoplasmic extracts, and 6-h and 24-h treatments reached significance; F(3,8) = 24.28, p = 0.0002. Conversely, morphine did not significantly alter FHC expression in nuclear extracts at any time; F(3,8) = 1.644, p = 0.2549. Both experiments were analyzed by one-way ANOVA and Dunnett post hoc.