nt. J. Mol. Sci. 2021, 22,Figure Recombinant sea bass AmhHis6 and and His6 Amh induce form two Amh receptor Figure two.2. Recombinant sea bass AmhHis6His6Amh induce sea bass sea bass variety two Amh receptor (Amhr2)-dependent BRE-luc reporter activity. African green green monkey kidney fibroblast-like (COS-7) (Amhr2)-dependent BRE-luc reporter activity. African monkey kidney fibroblast-like (COS7) cells weretransientlytransfectedAmhHis6 bass Amhr2, along withsea Bre-Luc reporter and pRL- pRL-TK Figure 2. Recombinant transfected with sea bass Amhr2, in addition to the type 2 Amh receptor cells were transiently sea bass with sea and His6Amh induce the bass Bre-Luc reporter and TK plasmids. The cells were incubated activity. African green monkey kidney fibroblast-like (COS(Amhr2)-dependent BRE-luc reporter with three diverse concentrations of purified recombinant plasmids. The 6cells were incubated with FBS culture three various concentrations of purified recombinant sea sea cells had been transiently6transfected withasea bass Amhr2,medium. Firefly luciferase activities had been 7) bass AmhHis or His Amh for 24 h in 1 in conjunction with the Bre-Luc reporter and pRLbass AmhHis6 or His6 Amh for 24 h in athreeFBS culture medium. induction of recombinant 1 various concentrationsFirefly luciferase activities had been normalized for the cells Renilla luciferase. The results shown are the fold of purified normalized TK plasmids. pRL-TK were incubated with luciferaseAmhHispRL-TK Renilla 24 h inside a 1 FBS(RLUs), of shownFirefly luciferase values of cells normalized to or His6Amh relative light units culture medium. will be the fold induction have been sea bass activity,6expressed as for luciferase. The outcomes every single therapy over the activities of normalized treated withactivity,media obtainedrelative pastoris expressingare of each and every pPIC9K vector. Data repnormalized culture expressedluciferase.P. light units (RLUs), the foldCathepsin B Inhibitor supplier treatment over the values of cells luciferase to pRL-TK Renilla as from the final results shown the empty induction of normalized resent the imply SEM of 3 independent Caspase 2 Inhibitor MedChemExpress experiments (in triplicate for each and every condition) and had been luciferase activity, expressed as obtained from P.(RLUs), ofexpressing theover the values of cells treated with culture media relative light units pastoris every treatment empty pPIC9K vector. Information analyzed by ANOVAmedia obtained from P. pastoris expressing the empty with asterisks are signiftreated with culture followed by Tukey’s considerable interaction test. Bars pPIC9K vector. Data reprepresent the SEM 3 independent experiments (in icantly unique mean0.05;threeof0.001) and ns indicates there were no fortriplicate for each situation) and resent the imply( p of p SEM independent experiments (in triplicate considerable differences. have been every condition) and have been analyzed by ANOVA followed by Tukey’s interaction test. Bars withtest. Barsare signifanalyzed by ANOVA followed by Tukey’s substantial important interaction asterisks with asterisks are substantially diverse ( 0.05; and ns indicates there were there had been variations. icantly unique ( p 0.05;p p 0.001) p 0.001) and ns indicatesno significantno significant differences.Figure 3. Human AMH induces sea bass Amhr2-dependent BRE-luc reporter activity. COS-7 cells have been transiently transfected with sea bass Amhr2, together with the BRE-Luc reporter and pRL-TK plasmids. The cells were incubated with three distinct concentrations of hAMH for 24 h within a 1 FBS culture medium. Firefly l