Fig. S1 p60TRP regulates endocytic recycling of the γ-opioid receptor (Oprd1). NSCs were grown as described in 'Materials and Methods' for 4P before stimulated for nine days with the specific Oprd1 agonist SNC80 (10 nM). p60TRP-transfected NSCs show clearly less Oprd1 expression before and upon stimulation with SNC80. Quantitative analysis of representative Western-blots is shown (* = P < 0.05, compared with control (Mock/Gfp-transfected)).

Fig. S2 Over-expression of p60TRP in NSCs and PC12 cells enhances the ATP metabolism. Stable PC12 cell lines over-expressing (+p60) and knock-down (-p60) p60TRP were established as described in 'Materials and Methods' using a lentivirus-based transfection system. Controls (C) were mock/Gfp-transfected cells. Tubulin (Tuba1a) was used as loading control (A). Comparison of the expression and cleavage pattern of p60TRP in NSCs and PC12 cells. NSCs were grown as described in supplemental 'Materials and Methods' before subjected to cell lyses along with PC12 cells and Western blot analyses were performed. Interestingly, only the proliferating NSCs show the specific p60TRP-cleaved 35 kDa band (as in Fig. 1A) (B). p60TRP exists as homo-dimer. p60TRP-expressing NSCs were lysed and treated with 1% DTT (dithiothreitol), 0.05% Nonidet P40 (NP40) and 1% 3-{(3-Cholamidopropyl)dimethylammonio}-1-propanesulfonate (CHAPS) to completely denature the nonionic interactions and disulfide bridges in p60TRP-dimers. 1 = control, 2 = DTT-and other non-ionic detergent-treated lysate (C). P60TRP-over-expressing PC12 cells show a higher ATP metabolism ratio than control cells while p60TRP-knock-down remained unchanged (D). p60TRP-transfected NSCs show a higher ATP metabolism ratio than control cells. ATP metabolism assays were performed as described in supplemental 'Materials and Methods', four times in duplicates. Quantification in (D) and (E) is represented as the mean (± SD) of four independent determinations, each performed in triplicate (*P < 0.05, compared with control cells) (E).

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