cleaved the pro-peptide in vivo, creating a mature sea that was observed involving pPICK9-His6 Amh and pPICK9-AmhHis6 constructs, indicating bass AmhC of 125position with the was -tag had tiny impact around the expressionas detected bassWest-(data the kDa, which His6 secreted in to the culture media, levels of sea by Amh not shown). ern blot following purification (Figure 1C). The sea bass His6Amh and AmhHis6 proteins wereslightly smaller sized than that previously obtained making use of CHO cells [30] and their relative sizes two.two. Functional Characterization of Sea Bass Amh also differed involving each (Figure 1C). In addition, tiny difference in expression levels To test the bioactivity of recombinant sea bass His6 Amh and AmhHis6 proteins, we was observed between pPICK9-His6Amh and pPICK9-AmhHisbass Amhr2 activity [30]. Each applied a cell-based reporter assay that depends on sea six constructs, indicating that the positionrecombinant6-tagbass Amh have been able to improve the activity of your CCR5 Antagonist Formulation BRE-Luc reporter of your His sea had small effect around the expression levels of sea bass Amh (data not shown). inside a dose-dependent manner (Figure two). Thinking about the slightly larger production genescale of AmhHis6 with respect to His6 Amh, only the former was utilised within the explant culture experiments. In a preceding study we demonstrated the capacity of sea bass AmhInt. J. Mol. Sci. 2021, 22, x FOR PEER REVIEW4 of2.two. Functional Characterization of Sea Bass Amh To test the bioactivity of recombinant sea bass His6Amh and AmhHis6 proteins, we two.2. a cell-based reporter assay Sea depends usedFunctional Characterization of thatBass Amh on sea bass Amhr2 activity [30]. Both re4 of 19 combinant sea bass Amh wererecombinant sea bassactivity in the BRE-Luc reporter gene To test the bioactivity of capable to improve the His6Amh and AmhHis6 proteins, we inside a dose-dependent manner (Figure 2). Taking into consideration the slightly larger production scale made use of a cell-based reporter assay that depends upon sea bass Amhr2 activity [30]. Both reof AmhHis6sea bass Amh to Hisable to only the the activity on the BRE-Luc reporter gene combinant with respect have been 6Amh, enhance former was utilized within the explant culture experiments. Within a prior study we demonstrated the capacity of sea bass Amh to actiin a dose-dependent manner (Figure two). Thinking about the slightly Dopamine Receptor Agonist Accession greater production scale to activate with respect and, to finish the former was used in the explant culture vate human 6human AMHR2 [30], and, to this interspecies functional comparison, the of AmhHis AMHR2 [30], to His6Amh, only complete this interspecies functional comparison, the ability of human to study the sea bass Amhr2 was tested. tested. The to actiability of humanaAMH AMH to activate the sea bass Amhr2 wasThe benefits showed showed experiments. In previousactivatewe demonstrated the capacity of sea bass Amh benefits drastically greater [30], and, activity than that obtained functional comparison, the drastically larger luciferase activity than that obtained with thethe controlall all tested doses, vate human AMHR2 luciferase to complete this interspecies with control at at tested along with a of a dose-response activate observed for AmhHis6 (Figure The doses, and human AMH to trend, observed bassAmhHis6 (Figure 3). capacity dose-response trend, as because the sea for Amhr2 was tested.three). outcomes showed drastically higher luciferase activity than that obtained using the handle at all tested doses, and a dose-response trend, as observed for AmhHis6 (Figure 3).I